anno_start anno_end anno_text entity_type sentence section 24 34 Coenzyme A chemical The Structural Basis of Coenzyme A Recycling in a Bacterial Organelle TITLE 50 59 Bacterial taxonomy_domain The Structural Basis of Coenzyme A Recycling in a Bacterial Organelle TITLE 0 9 Bacterial taxonomy_domain Bacterial Microcompartments (BMCs) are proteinaceous organelles that encapsulate critical segments of autotrophic and heterotrophic metabolic pathways; they are functionally diverse and are found across 23 different phyla. ABSTRACT 10 27 Microcompartments complex_assembly Bacterial Microcompartments (BMCs) are proteinaceous organelles that encapsulate critical segments of autotrophic and heterotrophic metabolic pathways; they are functionally diverse and are found across 23 different phyla. ABSTRACT 29 33 BMCs complex_assembly Bacterial Microcompartments (BMCs) are proteinaceous organelles that encapsulate critical segments of autotrophic and heterotrophic metabolic pathways; they are functionally diverse and are found across 23 different phyla. ABSTRACT 16 25 catabolic protein_state The majority of catabolic BMCs (metabolosomes) compartmentalize a common core of enzymes to metabolize compounds via a toxic and/or volatile aldehyde intermediate. ABSTRACT 26 30 BMCs complex_assembly The majority of catabolic BMCs (metabolosomes) compartmentalize a common core of enzymes to metabolize compounds via a toxic and/or volatile aldehyde intermediate. ABSTRACT 32 45 metabolosomes complex_assembly The majority of catabolic BMCs (metabolosomes) compartmentalize a common core of enzymes to metabolize compounds via a toxic and/or volatile aldehyde intermediate. ABSTRACT 141 149 aldehyde chemical The majority of catabolic BMCs (metabolosomes) compartmentalize a common core of enzymes to metabolize compounds via a toxic and/or volatile aldehyde intermediate. ABSTRACT 16 35 phosphotransacylase protein_type The core enzyme phosphotransacylase (PTAC) recycles Coenzyme A and generates an acyl phosphate that can serve as an energy source. ABSTRACT 37 41 PTAC protein_type The core enzyme phosphotransacylase (PTAC) recycles Coenzyme A and generates an acyl phosphate that can serve as an energy source. ABSTRACT 52 62 Coenzyme A chemical The core enzyme phosphotransacylase (PTAC) recycles Coenzyme A and generates an acyl phosphate that can serve as an energy source. ABSTRACT 80 94 acyl phosphate chemical The core enzyme phosphotransacylase (PTAC) recycles Coenzyme A and generates an acyl phosphate that can serve as an energy source. ABSTRACT 4 8 PTAC protein_type The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 39 52 metabolosomes complex_assembly The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 54 58 PduL protein_type The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 92 96 PTAC protein_type The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 114 135 fermentative bacteria taxonomy_domain The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 137 140 Pta protein_type The PTAC predominantly associated with metabolosomes (PduL) has no sequence homology to the PTAC ubiquitous among fermentative bacteria (Pta). ABSTRACT 36 40 PduL protein_type Here, we report two high-resolution PduL crystal structures with bound substrates. ABSTRACT 41 59 crystal structures evidence Here, we report two high-resolution PduL crystal structures with bound substrates. ABSTRACT 60 81 with bound substrates protein_state Here, we report two high-resolution PduL crystal structures with bound substrates. ABSTRACT 4 8 PduL protein_type The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 9 13 fold structure_element The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 30 34 that structure_element The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 38 41 Pta protein_type The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 57 76 dimetal active site site The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 137 149 housekeeping protein_state The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 150 154 PTAC protein_type The PduL fold is unrelated to that of Pta; it contains a dimetal active site involved in a catalytic mechanism distinct from that of the housekeeping PTAC. ABSTRACT 13 17 PduL protein_type Accordingly, PduL and Pta exemplify functional, but not structural, convergent evolution. ABSTRACT 22 25 Pta protein_type Accordingly, PduL and Pta exemplify functional, but not structural, convergent evolution. ABSTRACT 4 8 PduL protein_type The PduL structure, in the context of the catalytic core, completes our understanding of the structural basis of cofactor recycling in the metabolosome lumen. ABSTRACT 9 18 structure evidence The PduL structure, in the context of the catalytic core, completes our understanding of the structural basis of cofactor recycling in the metabolosome lumen. ABSTRACT 139 151 metabolosome complex_assembly The PduL structure, in the context of the catalytic core, completes our understanding of the structural basis of cofactor recycling in the metabolosome lumen. ABSTRACT 25 34 structure evidence This study describes the structure of a novel phosphotransacylase enzyme that facilitates the recycling of the essential cofactor acetyl-CoA within a bacterial organelle and discusses the properties of the enzyme's active site and how it is packaged into the organelle. ABSTRACT 46 65 phosphotransacylase protein_type This study describes the structure of a novel phosphotransacylase enzyme that facilitates the recycling of the essential cofactor acetyl-CoA within a bacterial organelle and discusses the properties of the enzyme's active site and how it is packaged into the organelle. ABSTRACT 130 140 acetyl-CoA chemical This study describes the structure of a novel phosphotransacylase enzyme that facilitates the recycling of the essential cofactor acetyl-CoA within a bacterial organelle and discusses the properties of the enzyme's active site and how it is packaged into the organelle. ABSTRACT 150 159 bacterial taxonomy_domain This study describes the structure of a novel phosphotransacylase enzyme that facilitates the recycling of the essential cofactor acetyl-CoA within a bacterial organelle and discusses the properties of the enzyme's active site and how it is packaged into the organelle. ABSTRACT 215 226 active site site This study describes the structure of a novel phosphotransacylase enzyme that facilitates the recycling of the essential cofactor acetyl-CoA within a bacterial organelle and discusses the properties of the enzyme's active site and how it is packaged into the organelle. ABSTRACT 57 60 ATP chemical In metabolism, molecules with “high-energy” bonds (e.g., ATP and Acetyl~CoA) are critical for both catabolic and anabolic processes. ABSTRACT 65 75 Acetyl~CoA chemical In metabolism, molecules with “high-energy” bonds (e.g., ATP and Acetyl~CoA) are critical for both catabolic and anabolic processes. ABSTRACT 4 23 phosphotransacylase protein_type The phosphotransacylase (Pta) enzyme catalyzes the conversion between acyl-CoA and acyl-phosphate. ABSTRACT 25 28 Pta protein_type The phosphotransacylase (Pta) enzyme catalyzes the conversion between acyl-CoA and acyl-phosphate. ABSTRACT 70 78 acyl-CoA chemical The phosphotransacylase (Pta) enzyme catalyzes the conversion between acyl-CoA and acyl-phosphate. ABSTRACT 83 97 acyl-phosphate chemical The phosphotransacylase (Pta) enzyme catalyzes the conversion between acyl-CoA and acyl-phosphate. ABSTRACT 32 40 acyl-CoA chemical This reaction directly links an acyl-CoA with ATP generation via substrate-level phosphorylation, producing short-chain fatty acids (e.g., acetate), and also provides a path for short-chain fatty acids to enter central metabolism. ABSTRACT 46 49 ATP chemical This reaction directly links an acyl-CoA with ATP generation via substrate-level phosphorylation, producing short-chain fatty acids (e.g., acetate), and also provides a path for short-chain fatty acids to enter central metabolism. ABSTRACT 108 131 short-chain fatty acids chemical This reaction directly links an acyl-CoA with ATP generation via substrate-level phosphorylation, producing short-chain fatty acids (e.g., acetate), and also provides a path for short-chain fatty acids to enter central metabolism. ABSTRACT 139 146 acetate chemical This reaction directly links an acyl-CoA with ATP generation via substrate-level phosphorylation, producing short-chain fatty acids (e.g., acetate), and also provides a path for short-chain fatty acids to enter central metabolism. ABSTRACT 178 201 short-chain fatty acids chemical This reaction directly links an acyl-CoA with ATP generation via substrate-level phosphorylation, producing short-chain fatty acids (e.g., acetate), and also provides a path for short-chain fatty acids to enter central metabolism. ABSTRACT 33 42 conserved protein_state Due to this key function, Pta is conserved across the bacterial kingdom. ABSTRACT 54 71 bacterial kingdom taxonomy_domain Due to this key function, Pta is conserved across the bacterial kingdom. ABSTRACT 24 43 phosphotransacylase protein_type Recently, a new type of phosphotransacylase was described that shares no evolutionary relation to Pta. ABSTRACT 98 101 Pta protein_type Recently, a new type of phosphotransacylase was described that shares no evolutionary relation to Pta. ABSTRACT 13 17 PduL protein_type This enzyme, PduL, is exclusively associated with organelles called bacterial microcompartments, which are used to catabolize various compounds. ABSTRACT 22 33 exclusively protein_state This enzyme, PduL, is exclusively associated with organelles called bacterial microcompartments, which are used to catabolize various compounds. ABSTRACT 68 77 bacterial taxonomy_domain This enzyme, PduL, is exclusively associated with organelles called bacterial microcompartments, which are used to catabolize various compounds. ABSTRACT 78 95 microcompartments complex_assembly This enzyme, PduL, is exclusively associated with organelles called bacterial microcompartments, which are used to catabolize various compounds. ABSTRACT 14 18 PduL protein_type Not only does PduL facilitate substrate level phosphorylation, but it also is critical for cofactor recycling within, and product efflux from, the organelle. ABSTRACT 3 9 solved experimental_method We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 14 23 structure evidence We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 32 42 convergent protein_state We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 43 62 phosphotransacylase protein_type We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 122 125 Pta protein_type We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 141 152 active site site We solved the structure of this convergent phosphotransacylase and show that it is completely structurally different from Pta, including its active site architecture. ABSTRACT 0 9 Bacterial taxonomy_domain Bacterial Microcompartments (BMCs) are organelles that encapsulate enzymes for sequential biochemical reactions within a protein shell. INTRO 10 27 Microcompartments complex_assembly Bacterial Microcompartments (BMCs) are organelles that encapsulate enzymes for sequential biochemical reactions within a protein shell. INTRO 29 33 BMCs complex_assembly Bacterial Microcompartments (BMCs) are organelles that encapsulate enzymes for sequential biochemical reactions within a protein shell. INTRO 129 134 shell structure_element Bacterial Microcompartments (BMCs) are organelles that encapsulate enzymes for sequential biochemical reactions within a protein shell. INTRO 4 9 shell structure_element The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 86 95 hexagonal protein_state The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 97 102 BMC-H complex_assembly The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 107 112 BMC-T complex_assembly The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 117 127 pentagonal protein_state The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 129 134 BMC-P complex_assembly The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 163 173 polyhedral protein_state The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 174 179 shell structure_element The shell is typically composed of three types of protein subunits, which form either hexagonal (BMC-H and BMC-T) or pentagonal (BMC-P) tiles that assemble into a polyhedral shell. INTRO 18 23 shell structure_element The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 50 58 hexamers oligomeric_state The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 92 97 pores site The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 109 125 highly conserved protein_state The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 127 132 polar protein_state The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 133 141 residues structure_element The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 219 224 shell structure_element The facets of the shell are composed primarily of hexamers that are typically perforated by pores lined with highly conserved, polar residues that presumably function as the conduits for metabolites into and out of the shell. INTRO 4 57 vitamin B12-dependent propanediol-utilizing (PDU) BMC complex_assembly The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 106 115 catabolic protein_state The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 116 120 BMCs complex_assembly The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 191 203 ethanolamine chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 205 212 choline chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 214 220 fucose chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 222 230 rhamnose chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 236 243 ethanol chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 276 284 aldehyde chemical The vitamin B12-dependent propanediol-utilizing (PDU) BMC was one of the first functionally characterized catabolic BMCs; subsequently, other types have been implicated in the degradation of ethanolamine, choline, fucose, rhamnose, and ethanol, all of which produce different aldehyde intermediates (Table 1). INTRO 15 36 bioinformatic studies experimental_method More recently, bioinformatic studies have demonstrated the widespread distribution of BMCs among diverse bacterial phyla and grouped them into 23 different functional types. INTRO 86 90 BMCs complex_assembly More recently, bioinformatic studies have demonstrated the widespread distribution of BMCs among diverse bacterial phyla and grouped them into 23 different functional types. INTRO 105 120 bacterial phyla taxonomy_domain More recently, bioinformatic studies have demonstrated the widespread distribution of BMCs among diverse bacterial phyla and grouped them into 23 different functional types. INTRO 45 54 catabolic protein_state The reactions carried out in the majority of catabolic BMCs (also known as metabolosomes) fit a generalized biochemical paradigm for the oxidation of aldehydes (Fig 1). INTRO 55 59 BMCs complex_assembly The reactions carried out in the majority of catabolic BMCs (also known as metabolosomes) fit a generalized biochemical paradigm for the oxidation of aldehydes (Fig 1). INTRO 75 88 metabolosomes complex_assembly The reactions carried out in the majority of catabolic BMCs (also known as metabolosomes) fit a generalized biochemical paradigm for the oxidation of aldehydes (Fig 1). INTRO 150 159 aldehydes chemical The reactions carried out in the majority of catabolic BMCs (also known as metabolosomes) fit a generalized biochemical paradigm for the oxidation of aldehydes (Fig 1). INTRO 16 19 BMC complex_assembly This involves a BMC-encapsulated signature enzyme that generates a toxic and/or volatile aldehyde that the BMC shell sequesters from the cytosol. INTRO 89 97 aldehyde chemical This involves a BMC-encapsulated signature enzyme that generates a toxic and/or volatile aldehyde that the BMC shell sequesters from the cytosol. INTRO 107 110 BMC complex_assembly This involves a BMC-encapsulated signature enzyme that generates a toxic and/or volatile aldehyde that the BMC shell sequesters from the cytosol. INTRO 111 116 shell structure_element This involves a BMC-encapsulated signature enzyme that generates a toxic and/or volatile aldehyde that the BMC shell sequesters from the cytosol. INTRO 4 12 aldehyde chemical The aldehyde is subsequently converted into an acyl-CoA by aldehyde dehydrogenase, which uses NAD+ and CoA as cofactors. INTRO 47 55 acyl-CoA chemical The aldehyde is subsequently converted into an acyl-CoA by aldehyde dehydrogenase, which uses NAD+ and CoA as cofactors. INTRO 59 81 aldehyde dehydrogenase protein_type The aldehyde is subsequently converted into an acyl-CoA by aldehyde dehydrogenase, which uses NAD+ and CoA as cofactors. INTRO 94 98 NAD+ chemical The aldehyde is subsequently converted into an acyl-CoA by aldehyde dehydrogenase, which uses NAD+ and CoA as cofactors. INTRO 103 106 CoA chemical The aldehyde is subsequently converted into an acyl-CoA by aldehyde dehydrogenase, which uses NAD+ and CoA as cofactors. INTRO 73 86 protein shell structure_element These two cofactors are relatively large, and their diffusion across the protein shell is thought to be restricted, necessitating their regeneration within the BMC lumen. INTRO 160 163 BMC complex_assembly These two cofactors are relatively large, and their diffusion across the protein shell is thought to be restricted, necessitating their regeneration within the BMC lumen. INTRO 0 4 NAD+ chemical NAD+ is recycled via alcohol dehydrogenase, and CoA is recycled via phosphotransacetylase (PTAC) (Fig 1). INTRO 21 42 alcohol dehydrogenase protein_type NAD+ is recycled via alcohol dehydrogenase, and CoA is recycled via phosphotransacetylase (PTAC) (Fig 1). INTRO 48 51 CoA chemical NAD+ is recycled via alcohol dehydrogenase, and CoA is recycled via phosphotransacetylase (PTAC) (Fig 1). INTRO 68 89 phosphotransacetylase protein_type NAD+ is recycled via alcohol dehydrogenase, and CoA is recycled via phosphotransacetylase (PTAC) (Fig 1). INTRO 91 95 PTAC protein_type NAD+ is recycled via alcohol dehydrogenase, and CoA is recycled via phosphotransacetylase (PTAC) (Fig 1). INTRO 25 28 BMC complex_assembly The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 33 47 acyl-phosphate chemical The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 78 81 ATP chemical The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 86 97 acyl kinase protein_type The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 117 125 acyl-CoA chemical The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 129 132 Pta protein_type The final product of the BMC, an acyl-phosphate, can then be used to generate ATP via acyl kinase, or revert back to acyl-CoA by Pta for biosynthesis. INTRO 18 53 aldehyde and alcohol dehydrogenases protein_type Collectively, the aldehyde and alcohol dehydrogenases, as well as the PTAC, constitute the common metabolosome core. INTRO 70 74 PTAC protein_type Collectively, the aldehyde and alcohol dehydrogenases, as well as the PTAC, constitute the common metabolosome core. INTRO 98 110 metabolosome complex_assembly Collectively, the aldehyde and alcohol dehydrogenases, as well as the PTAC, constitute the common metabolosome core. INTRO 29 47 aldehyde-degrading protein_state General biochemical model of aldehyde-degrading BMCs (metabolosomes) illustrating the common metabolosome core enzymes and reactions. FIG 48 52 BMCs complex_assembly General biochemical model of aldehyde-degrading BMCs (metabolosomes) illustrating the common metabolosome core enzymes and reactions. FIG 54 67 metabolosomes complex_assembly General biochemical model of aldehyde-degrading BMCs (metabolosomes) illustrating the common metabolosome core enzymes and reactions. FIG 93 105 metabolosome complex_assembly General biochemical model of aldehyde-degrading BMCs (metabolosomes) illustrating the common metabolosome core enzymes and reactions. FIG 39 43 PTAC protein_type Substrates and cofactors involving the PTAC reaction are shown in red; other substrates and enzymes are shown in black, and other cofactors are shown in gray. FIG 28 37 catabolic protein_state Characterized and predicted catabolic BMC (metabolosome) types that represent the aldehyde-degrading paradigm (for definition of types see Kerfeld and Erbilgin). TABLE 38 41 BMC complex_assembly Characterized and predicted catabolic BMC (metabolosome) types that represent the aldehyde-degrading paradigm (for definition of types see Kerfeld and Erbilgin). TABLE 43 55 metabolosome complex_assembly Characterized and predicted catabolic BMC (metabolosome) types that represent the aldehyde-degrading paradigm (for definition of types see Kerfeld and Erbilgin). TABLE 82 90 aldehyde chemical Characterized and predicted catabolic BMC (metabolosome) types that represent the aldehyde-degrading paradigm (for definition of types see Kerfeld and Erbilgin). TABLE 5 9 PTAC protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 27 35 Aldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 38 41 PDU complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 43 47 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 48 63 propionaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 66 70 EUT1 complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 71 78 PTA_PTB protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 79 91 acetaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 94 98 EUT2 complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 99 103 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 104 116 acetaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 119 122 ETU complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 128 140 acetaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 143 151 GRM1/CUT complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 152 156 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 157 169 acetaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 172 176 GRM2 complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 177 181 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 182 194 acetaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 197 204 GRM3*,4 complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 205 209 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 210 225 propionaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 228 236 GRM5/GRP complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 237 241 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 242 257 propionaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 260 263 PVM complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 265 269 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 270 282 lactaldehyde chemical "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 285 291 RMM1,2 complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 307 310 SPU complex_assembly "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 311 315 PduL protein_type "Name PTAC Type Sequestered Aldehyde PDU* PduL propionaldehyde EUT1 PTA_PTB acetaldehyde EUT2 PduL acetaldehyde ETU None acetaldehyde GRM1/CUT PduL acetaldehyde GRM2 PduL acetaldehyde GRM3*,4 PduL propionaldehyde GRM5/GRP PduL propionaldehyde PVM* PduL lactaldehyde RMM1,2 None unknown SPU PduL unknown " TABLE 2 6 PduL protein_type * PduL from these functional types of metabolosomes were purified in this study. TABLE 38 51 metabolosomes complex_assembly * PduL from these functional types of metabolosomes were purified in this study. TABLE 51 54 BMC complex_assembly The activities of core enzymes are not confined to BMC-associated functions: aldehyde and alcohol dehydrogenases are utilized in diverse metabolic reactions, and PTAC catalyzes a key biochemical reaction in the process of obtaining energy during fermentation. INTRO 77 112 aldehyde and alcohol dehydrogenases protein_type The activities of core enzymes are not confined to BMC-associated functions: aldehyde and alcohol dehydrogenases are utilized in diverse metabolic reactions, and PTAC catalyzes a key biochemical reaction in the process of obtaining energy during fermentation. INTRO 162 166 PTAC protein_type The activities of core enzymes are not confined to BMC-associated functions: aldehyde and alcohol dehydrogenases are utilized in diverse metabolic reactions, and PTAC catalyzes a key biochemical reaction in the process of obtaining energy during fermentation. INTRO 31 35 PTAC protein_type The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 43 57 acetate kinase protein_type The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 59 62 Ack protein_type The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 79 82 ATP chemical The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 117 125 pyruvate chemical The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 129 136 acetate chemical The concerted functioning of a PTAC and an acetate kinase (Ack) is crucial for ATP generation in the fermentation of pyruvate to acetate (see Reactions 1 and 2). INTRO 45 67 fermentative organisms taxonomy_domain Both enzymes are, however, not restricted to fermentative organisms. INTRO 56 63 acetate chemical They can also work in the reverse direction to activate acetate to the CoA-thioester. INTRO 71 84 CoA-thioester chemical They can also work in the reverse direction to activate acetate to the CoA-thioester. INTRO 68 76 archaeal taxonomy_domain This occurs, for example, during acetoclastic methanogenesis in the archaeal Methanosarcina species. INTRO 77 99 Methanosarcina species taxonomy_domain This occurs, for example, during acetoclastic methanogenesis in the archaeal Methanosarcina species. INTRO 13 25 acetyl-S-CoA chemical Reaction 1: acetyl-S-CoA + Pi ←→ acetyl phosphate + CoA-SH (PTAC) INTRO 28 30 Pi chemical Reaction 1: acetyl-S-CoA + Pi ←→ acetyl phosphate + CoA-SH (PTAC) INTRO 34 50 acetyl phosphate chemical Reaction 1: acetyl-S-CoA + Pi ←→ acetyl phosphate + CoA-SH (PTAC) INTRO 53 59 CoA-SH chemical Reaction 1: acetyl-S-CoA + Pi ←→ acetyl phosphate + CoA-SH (PTAC) INTRO 61 65 PTAC protein_type Reaction 1: acetyl-S-CoA + Pi ←→ acetyl phosphate + CoA-SH (PTAC) INTRO 13 29 acetyl phosphate chemical Reaction 2: acetyl phosphate + ADP ←→ acetate + ATP (Ack) INTRO 32 35 ADP chemical Reaction 2: acetyl phosphate + ADP ←→ acetate + ATP (Ack) INTRO 39 46 acetate chemical Reaction 2: acetyl phosphate + ADP ←→ acetate + ATP (Ack) INTRO 49 52 ATP chemical Reaction 2: acetyl phosphate + ADP ←→ acetate + ATP (Ack) INTRO 54 57 Ack protein_type Reaction 2: acetyl phosphate + ADP ←→ acetate + ATP (Ack) INTRO 14 18 PTAC protein_type The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 20 23 Pta protein_type The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 60 70 eukaryotes taxonomy_domain The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 75 82 archaea taxonomy_domain The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 109 117 bacteria taxonomy_domain The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 130 139 bacterial taxonomy_domain The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 221 248 PTA_PTB phosphotransacylase protein_type The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 262 269 PF01515 structure_element The canonical PTAC, Pta, is an ancient enzyme found in some eukaryotes and archaea, and widespread among the bacteria; 90% of the bacterial genomes in the Integrated Microbial Genomes database contain a gene encoding the PTA_PTB phosphotransacylase (Pfam domain PF01515). INTRO 0 3 Pta protein_type Pta has been extensively characterized due to its key role in fermentation. INTRO 32 36 PTAC protein_type More recently, a second type of PTAC without any sequence homology to Pta was identified. INTRO 70 73 Pta protein_type More recently, a second type of PTAC without any sequence homology to Pta was identified. INTRO 14 18 PduL protein_type This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 32 39 PF06130 structure_element This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 82 95 propionyl-CoA chemical This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 99 118 propionyl-phosphate chemical This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 144 147 BMC complex_assembly This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 189 196 PDU BMC complex_assembly This protein, PduL (Pfam domain PF06130), was shown to catalyze the conversion of propionyl-CoA to propionyl-phosphate and is associated with a BMC involved in propanediol utilization, the PDU BMC. INTRO 5 9 pduL gene Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 14 17 pta gene Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 78 82 BMCs complex_assembly Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 97 101 PduL protein_type Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 166 184 metabolosome locus gene Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 186 190 EUT1 gene Both pduL and pta genes can be found in genetic loci of functionally distinct BMCs, although the PduL type is much more prevalent, being found in all but one type of metabolosome locus: EUT1 (Table 1). INTRO 86 93 PF06130 structure_element Furthermore, in the Integrated Microbial Genomes Database, 91% of genomes that encode PF06130 also encode genes for shell proteins. INTRO 70 88 aldehyde-oxidizing protein_state As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 89 102 metabolosomes complex_assembly As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 104 108 PduL protein_type As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 197 200 Pta protein_type As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 246 256 acetyl-CoA chemical As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 258 262 PduL protein_type As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 272 279 PDU BMC complex_assembly As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 283 302 Salmonella enterica species As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 310 323 propionyl-CoA chemical As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 329 339 acetyl-CoA chemical As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 363 367 PduL protein_type As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 402 406 BMCs complex_assembly As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 450 453 CoA chemical As a member of the core biochemical machinery of functionally diverse aldehyde-oxidizing metabolosomes, PduL must have a certain level of substrate plasticity (see Table 1) that is not required of Pta, which has generally been observed to prefer acetyl-CoA. PduL from the PDU BMC of Salmonella enterica favors propionyl-CoA over acetyl-CoA, and it is likely that PduL orthologs in functionally diverse BMCs would have substrate preferences for other CoA derivatives. INTRO 31 45 BMC-associated protein_state Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 46 50 PduL protein_type Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 77 98 encapsulation peptide structure_element Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 100 102 EP structure_element Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 166 169 BMC complex_assembly Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 170 175 shell structure_element Another distinctive feature of BMC-associated PduL homologs is an N-terminal encapsulation peptide (EP) that is thought to “target” proteins for encapsulation by the BMC shell. INTRO 0 3 EPs structure_element EPs are frequently found on BMC-associated proteins and have been shown to interact with shell proteins. INTRO 28 51 BMC-associated proteins protein_type EPs are frequently found on BMC-associated proteins and have been shown to interact with shell proteins. INTRO 0 3 EPs structure_element EPs have also been observed to cause proteins to aggregate, and this has recently been suggested to be functionally relevant as an initial step in metabolosome assembly, in which a multifunctional protein core is formed, around which the shell assembles. INTRO 147 159 metabolosome complex_assembly EPs have also been observed to cause proteins to aggregate, and this has recently been suggested to be functionally relevant as an initial step in metabolosome assembly, in which a multifunctional protein core is formed, around which the shell assembles. INTRO 238 243 shell structure_element EPs have also been observed to cause proteins to aggregate, and this has recently been suggested to be functionally relevant as an initial step in metabolosome assembly, in which a multifunctional protein core is formed, around which the shell assembles. INTRO 20 32 metabolosome complex_assembly Of the three common metabolosome core enzymes, crystal structures are available for both the alcohol and aldehyde dehydrogenases. INTRO 47 65 crystal structures evidence Of the three common metabolosome core enzymes, crystal structures are available for both the alcohol and aldehyde dehydrogenases. INTRO 93 128 alcohol and aldehyde dehydrogenases protein_type Of the three common metabolosome core enzymes, crystal structures are available for both the alcohol and aldehyde dehydrogenases. INTRO 17 26 structure evidence In contrast, the structure of PduL, the PTAC found in the vast majority of catabolic BMCs, has not been determined. INTRO 30 34 PduL protein_type In contrast, the structure of PduL, the PTAC found in the vast majority of catabolic BMCs, has not been determined. INTRO 40 44 PTAC protein_type In contrast, the structure of PduL, the PTAC found in the vast majority of catabolic BMCs, has not been determined. INTRO 75 84 catabolic protein_state In contrast, the structure of PduL, the PTAC found in the vast majority of catabolic BMCs, has not been determined. INTRO 85 89 BMCs complex_assembly In contrast, the structure of PduL, the PTAC found in the vast majority of catabolic BMCs, has not been determined. INTRO 44 56 metabolosome complex_assembly This is a major gap in our understanding of metabolosome-encapsulated biochemistry and cofactor recycling. INTRO 66 69 BMC complex_assembly Moreover, it will be useful for guiding efforts to engineer novel BMC cores for biotechnological applications. INTRO 25 29 PduL protein_type The primary structure of PduL homologs is subdivided into two PF06130 domains, each roughly 80 residues in length. INTRO 62 69 PF06130 structure_element The primary structure of PduL homologs is subdivided into two PF06130 domains, each roughly 80 residues in length. INTRO 92 113 80 residues in length residue_range The primary structure of PduL homologs is subdivided into two PF06130 domains, each roughly 80 residues in length. INTRO 44 51 PF06130 structure_element No available protein structures contain the PF06130 domain, and homology searches using the primary structure of PduL do not return any significant results that would allow prediction of the structure. INTRO 64 81 homology searches experimental_method No available protein structures contain the PF06130 domain, and homology searches using the primary structure of PduL do not return any significant results that would allow prediction of the structure. INTRO 113 117 PduL protein_type No available protein structures contain the PF06130 domain, and homology searches using the primary structure of PduL do not return any significant results that would allow prediction of the structure. INTRO 191 200 structure evidence No available protein structures contain the PF06130 domain, and homology searches using the primary structure of PduL do not return any significant results that would allow prediction of the structure. INTRO 33 37 PduL protein_type Moreover, the evident novelty of PduL makes its structure interesting in the context of convergent evolution of PTAC function; to-date, only the Pta active site and catalytic mechanism is known. INTRO 48 57 structure evidence Moreover, the evident novelty of PduL makes its structure interesting in the context of convergent evolution of PTAC function; to-date, only the Pta active site and catalytic mechanism is known. INTRO 112 116 PTAC protein_type Moreover, the evident novelty of PduL makes its structure interesting in the context of convergent evolution of PTAC function; to-date, only the Pta active site and catalytic mechanism is known. INTRO 145 148 Pta protein_type Moreover, the evident novelty of PduL makes its structure interesting in the context of convergent evolution of PTAC function; to-date, only the Pta active site and catalytic mechanism is known. INTRO 149 160 active site site Moreover, the evident novelty of PduL makes its structure interesting in the context of convergent evolution of PTAC function; to-date, only the Pta active site and catalytic mechanism is known. INTRO 31 49 crystal structures evidence Here we report high-resolution crystal structures of a PduL-type PTAC in both CoA- and phosphate-bound forms, completing our understanding of the structural basis of catalysis by the metabolosome common core enzymes. INTRO 55 69 PduL-type PTAC protein_type Here we report high-resolution crystal structures of a PduL-type PTAC in both CoA- and phosphate-bound forms, completing our understanding of the structural basis of catalysis by the metabolosome common core enzymes. INTRO 78 82 CoA- protein_state Here we report high-resolution crystal structures of a PduL-type PTAC in both CoA- and phosphate-bound forms, completing our understanding of the structural basis of catalysis by the metabolosome common core enzymes. INTRO 87 102 phosphate-bound protein_state Here we report high-resolution crystal structures of a PduL-type PTAC in both CoA- and phosphate-bound forms, completing our understanding of the structural basis of catalysis by the metabolosome common core enzymes. INTRO 183 195 metabolosome complex_assembly Here we report high-resolution crystal structures of a PduL-type PTAC in both CoA- and phosphate-bound forms, completing our understanding of the structural basis of catalysis by the metabolosome common core enzymes. INTRO 80 83 Pta protein_type We propose a catalytic mechanism analogous but yet distinct from the ubiquitous Pta enzyme, highlighting the functional convergence of two enzymes with completely different structures and metal requirements. INTRO 65 69 PduL protein_type We also investigate the quaternary structures of three different PduL homologs and situate our findings in the context of organelle biogenesis in functionally diverse BMCs. INTRO 167 171 BMCs complex_assembly We also investigate the quaternary structures of three different PduL homologs and situate our findings in the context of organelle biogenesis in functionally diverse BMCs. INTRO 0 23 Structure Determination experimental_method Structure Determination of PduL RESULTS 27 31 PduL protein_type Structure Determination of PduL RESULTS 3 34 cloned, expressed, and purified experimental_method We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 51 55 PduL protein_type We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 92 96 BMCs complex_assembly We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 130 139 pdu locus gene We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 143 170 S. enterica Typhimurium LT2 species We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 172 177 sPduL protein We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 212 221 pvm locus gene We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 225 249 Planctomyces limnophilus species We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 251 256 pPduL protein We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 272 282 grm3 locus gene We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 286 319 Rhodopseudomonas palustris BisB18 species We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 321 326 rPduL protein We cloned, expressed, and purified three different PduL homologs from functionally distinct BMCs (Table 1): from the well-studied pdu locus in S. enterica Typhimurium LT2 (sPduL), from the recently characterized pvm locus in Planctomyces limnophilus (pPduL), and from the grm3 locus in Rhodopseudomonas palustris BisB18 (rPduL). RESULTS 16 27 full-length protein_state While purifying full-length sPduL, we observed a tendency to aggregation as described previously, with a large fraction of the expressed protein found in the insoluble fraction in a white, cake-like pellet. RESULTS 28 33 sPduL protein While purifying full-length sPduL, we observed a tendency to aggregation as described previously, with a large fraction of the expressed protein found in the insoluble fraction in a white, cake-like pellet. RESULTS 18 26 removing experimental_method Remarkably, after removing the N-terminal putative EP (27 amino acids), most of the sPduLΔEP protein was in the soluble fraction upon cell lysis. RESULTS 51 53 EP structure_element Remarkably, after removing the N-terminal putative EP (27 amino acids), most of the sPduLΔEP protein was in the soluble fraction upon cell lysis. RESULTS 55 69 27 amino acids residue_range Remarkably, after removing the N-terminal putative EP (27 amino acids), most of the sPduLΔEP protein was in the soluble fraction upon cell lysis. RESULTS 84 92 sPduLΔEP mutant Remarkably, after removing the N-terminal putative EP (27 amino acids), most of the sPduLΔEP protein was in the soluble fraction upon cell lysis. RESULTS 52 57 pPduL protein Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 62 67 rPduL protein Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 83 95 EP-truncated protein_state Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 109 120 full-length protein_state Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 169 174 sPduL protein Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 212 218 active protein_state Similar differences in solubility were observed for pPduL and rPduL when comparing EP-truncated forms to the full-length protein, but none were quite as dramatic as for sPduL. We confirmed that all homologs were active (S1a and S1b Fig). RESULTS 41 69 diffraction-quality crystals evidence Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 73 78 rPduL protein Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 85 93 removing experimental_method Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 118 120 EP structure_element Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 122 136 33 amino acids residue_range Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 156 164 rPduLΔEP mutant Among these, we were only able to obtain diffraction-quality crystals of rPduL after removing the N-terminal putative EP (33 amino acids, also see Fig 2a) (rPduLΔEP). RESULTS 0 9 Truncated protein_state Truncated rPduLΔEP had comparable enzymatic activity to the full-length enzyme (S1a Fig). RESULTS 10 18 rPduLΔEP mutant Truncated rPduLΔEP had comparable enzymatic activity to the full-length enzyme (S1a Fig). RESULTS 60 71 full-length protein_state Truncated rPduLΔEP had comparable enzymatic activity to the full-length enzyme (S1a Fig). RESULTS 23 35 R. palustris species Structural overview of R. palustris PduL from the grm3 locus. FIG 36 40 PduL protein_type Structural overview of R. palustris PduL from the grm3 locus. FIG 50 60 grm3 locus gene Structural overview of R. palustris PduL from the grm3 locus. FIG 39 44 rPduL protein (a) Primary and secondary structure of rPduL (tubes represent α-helices, arrows β-sheets and dashed line residues disordered in the structure. FIG 62 71 α-helices structure_element (a) Primary and secondary structure of rPduL (tubes represent α-helices, arrows β-sheets and dashed line residues disordered in the structure. FIG 80 88 β-sheets structure_element (a) Primary and secondary structure of rPduL (tubes represent α-helices, arrows β-sheets and dashed line residues disordered in the structure. FIG 132 141 structure evidence (a) Primary and secondary structure of rPduL (tubes represent α-helices, arrows β-sheets and dashed line residues disordered in the structure. FIG 4 24 first 33 amino acids residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 95 97 EP structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 98 109 alpha helix structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 111 113 α0 structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 120 129 truncated protein_state The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 130 138 rPduLΔEP mutant The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 148 160 crystallized experimental_method The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 173 174 M residue_name The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 175 176 G residue_name The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 177 178 V residue_name The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 225 233 domain 1 structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 234 241 D36-N46 residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 242 251 Q155-C224 residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 259 273 loop insertion structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 274 281 G61-E81 residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 289 297 domain 2 structure_element The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 298 305 R47-F60 residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 306 314 E82-A154 residue_range The first 33 amino acids are present only in the wildtype construct and contains the predicted EP alpha helix, α0); the truncated rPduLΔEP that was crystallized begins with M-G-V. Coloring is according to structural domains (domain 1 D36-N46/Q155-C224, blue; loop insertion G61-E81, grey; domain 2 R47-F60/E82-A154, red). FIG 0 27 Metal coordination residues site Metal coordination residues are highlighted in light blue and CoA contacting residues in magenta, residues contacting the CoA of the other chain are also outlined. FIG 62 85 CoA contacting residues site Metal coordination residues are highlighted in light blue and CoA contacting residues in magenta, residues contacting the CoA of the other chain are also outlined. FIG 122 125 CoA chemical Metal coordination residues are highlighted in light blue and CoA contacting residues in magenta, residues contacting the CoA of the other chain are also outlined. FIG 34 43 structure evidence (b) Cartoon representation of the structure colored by domains and including secondary structure numbering. FIG 87 96 structure evidence (b) Cartoon representation of the structure colored by domains and including secondary structure numbering. FIG 0 10 Coenzyme A chemical Coenzyme A is shown in magenta sticks and Zinc (grey) as spheres. FIG 42 46 Zinc chemical Coenzyme A is shown in magenta sticks and Zinc (grey) as spheres. FIG 3 29 collected a native dataset experimental_method We collected a native dataset from rPduLΔEP crystals diffracting to a resolution of 1.54 Å (Table 2). RESULTS 35 43 rPduLΔEP mutant We collected a native dataset from rPduLΔEP crystals diffracting to a resolution of 1.54 Å (Table 2). RESULTS 44 52 crystals evidence We collected a native dataset from rPduLΔEP crystals diffracting to a resolution of 1.54 Å (Table 2). RESULTS 8 34 mercury-derivative crystal experimental_method Using a mercury-derivative crystal form diffracting to 1.99 Å (Table 2), we obtained high quality electron density for model building and used the initial model to refine against the native data to Rwork/Rfree values of 18.9/22.1%. RESULTS 98 114 electron density evidence Using a mercury-derivative crystal form diffracting to 1.99 Å (Table 2), we obtained high quality electron density for model building and used the initial model to refine against the native data to Rwork/Rfree values of 18.9/22.1%. RESULTS 198 203 Rwork evidence Using a mercury-derivative crystal form diffracting to 1.99 Å (Table 2), we obtained high quality electron density for model building and used the initial model to refine against the native data to Rwork/Rfree values of 18.9/22.1%. RESULTS 204 209 Rfree evidence Using a mercury-derivative crystal form diffracting to 1.99 Å (Table 2), we obtained high quality electron density for model building and used the initial model to refine against the native data to Rwork/Rfree values of 18.9/22.1%. RESULTS 14 18 PduL protein_type There are two PduL molecules in the asymmetric unit of the P212121 unit cell. RESULTS 52 59 PduLΔEP mutant We were able to fit all of the primary structure of PduLΔEP into the electron density with the exception of three amino acids at the N-terminus and two amino acids at the C-terminus (Fig 2a); the model is of excellent quality (Table 2). RESULTS 69 85 electron density evidence We were able to fit all of the primary structure of PduLΔEP into the electron density with the exception of three amino acids at the N-terminus and two amino acids at the C-terminus (Fig 2a); the model is of excellent quality (Table 2). RESULTS 2 5 CoA chemical A CoA cofactor as well as two metal ions are clearly resolved in the density (for omit maps of CoA see S2 Fig). RESULTS 69 76 density evidence A CoA cofactor as well as two metal ions are clearly resolved in the density (for omit maps of CoA see S2 Fig). RESULTS 82 91 omit maps evidence A CoA cofactor as well as two metal ions are clearly resolved in the density (for omit maps of CoA see S2 Fig). RESULTS 95 98 CoA chemical A CoA cofactor as well as two metal ions are clearly resolved in the density (for omit maps of CoA see S2 Fig). RESULTS 14 18 PduL protein_type Structurally, PduL consists of two domains (Fig 2, blue/red), each a beta-barrel that is capped on both ends by short α-helices. RESULTS 35 42 domains structure_element Structurally, PduL consists of two domains (Fig 2, blue/red), each a beta-barrel that is capped on both ends by short α-helices. RESULTS 69 80 beta-barrel structure_element Structurally, PduL consists of two domains (Fig 2, blue/red), each a beta-barrel that is capped on both ends by short α-helices. RESULTS 118 127 α-helices structure_element Structurally, PduL consists of two domains (Fig 2, blue/red), each a beta-barrel that is capped on both ends by short α-helices. RESULTS 0 10 β-Barrel 1 structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 38 46 β strand structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 51 60 β strands structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 70 85 C-terminal half structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 112 114 β1 structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 116 123 β10-β14 structure_element β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 134 139 37–46 residue_range β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 144 151 155–224 residue_range β-Barrel 1 consists of the N-terminal β strand and β strands from the C-terminal half of the polypeptide chain (β1, β10-β14; residues 37–46 and 155–224). RESULTS 0 10 β-Barrel 2 structure_element β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 72 74 β2 structure_element β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 76 81 β5–β9 structure_element β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 92 97 47–60 residue_range β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 102 108 82–154 residue_range β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 148 175 short two-strand beta sheet structure_element β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 177 182 β3-β4 structure_element β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 193 198 61–81 residue_range β-Barrel 2 consists mainly of the central segment of primary structure (β2, β5–β9; residues 47–60 and 82–154) (Fig 2, red), but is interrupted by a short two-strand beta sheet (β3-β4, residues 61–81). RESULTS 5 12 β-sheet structure_element This β-sheet is involved in contacts between the two domains and forms a lid over the active site. RESULTS 86 97 active site site This β-sheet is involved in contacts between the two domains and forms a lid over the active site. RESULTS 25 30 Gln42 residue_name_number Residues in this region (Gln42, Pro43, Gly44), covering the active site, are strongly conserved (Fig 3). RESULTS 32 37 Pro43 residue_name_number Residues in this region (Gln42, Pro43, Gly44), covering the active site, are strongly conserved (Fig 3). RESULTS 39 44 Gly44 residue_name_number Residues in this region (Gln42, Pro43, Gly44), covering the active site, are strongly conserved (Fig 3). RESULTS 60 71 active site site Residues in this region (Gln42, Pro43, Gly44), covering the active site, are strongly conserved (Fig 3). RESULTS 77 95 strongly conserved protein_state Residues in this region (Gln42, Pro43, Gly44), covering the active site, are strongly conserved (Fig 3). RESULTS 82 85 Pta protein_type This structural arrangement is completely different from the functionally related Pta, which is composed of two domains, each consisting of a central flat beta sheet with alpha-helices on the top and bottom. RESULTS 112 119 domains structure_element This structural arrangement is completely different from the functionally related Pta, which is composed of two domains, each consisting of a central flat beta sheet with alpha-helices on the top and bottom. RESULTS 155 165 beta sheet structure_element This structural arrangement is completely different from the functionally related Pta, which is composed of two domains, each consisting of a central flat beta sheet with alpha-helices on the top and bottom. RESULTS 171 184 alpha-helices structure_element This structural arrangement is completely different from the functionally related Pta, which is composed of two domains, each consisting of a central flat beta sheet with alpha-helices on the top and bottom. RESULTS 38 42 PduL protein_type Primary structure conservation of the PduL protein family. FIG 34 61 multiple sequence alignment experimental_method Sequence logo calculated from the multiple sequence alignment of PduL homologs (see Materials and Methods), but not including putative EP sequences. FIG 65 69 PduL protein_type Sequence logo calculated from the multiple sequence alignment of PduL homologs (see Materials and Methods), but not including putative EP sequences. FIG 112 125 not including protein_state Sequence logo calculated from the multiple sequence alignment of PduL homologs (see Materials and Methods), but not including putative EP sequences. FIG 135 137 EP structure_element Sequence logo calculated from the multiple sequence alignment of PduL homologs (see Materials and Methods), but not including putative EP sequences. FIG 35 39 PduL protein_type Residues 100% conserved across all PduL homologs in our dataset are noted with an asterisk, and residues conserved in over 90% of sequences are noted with a colon. FIG 30 37 PF06130 structure_element The sequences aligning to the PF06130 domain (determined by BLAST) are highlighted in red and blue. FIG 66 71 rPduL protein The position numbers shown correspond to the residue numbering of rPduL; note that some positions in the logo represent gaps in the rPduL sequence. FIG 132 137 rPduL protein The position numbers shown correspond to the residue numbering of rPduL; note that some positions in the logo represent gaps in the rPduL sequence. FIG 14 18 PduL protein_type There are two PduL molecules in the asymmetric unit forming a butterfly-shaped dimer (Fig 4c). RESULTS 62 78 butterfly-shaped protein_state There are two PduL molecules in the asymmetric unit forming a butterfly-shaped dimer (Fig 4c). RESULTS 79 84 dimer oligomeric_state There are two PduL molecules in the asymmetric unit forming a butterfly-shaped dimer (Fig 4c). RESULTS 35 64 size exclusion chromatography experimental_method Consistent with this, results from size exclusion chromatography of rPduLΔEP suggest that it is a dimer in solution (Fig 5e). RESULTS 68 76 rPduLΔEP mutant Consistent with this, results from size exclusion chromatography of rPduLΔEP suggest that it is a dimer in solution (Fig 5e). RESULTS 98 103 dimer oligomeric_state Consistent with this, results from size exclusion chromatography of rPduLΔEP suggest that it is a dimer in solution (Fig 5e). RESULTS 4 13 interface site The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 55 62 monomer oligomeric_state The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 87 104 α-helices 2 and 4 structure_element The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 118 136 β-sheets 12 and 14 structure_element The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 171 178 adenine chemical The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 189 192 CoA chemical The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 198 204 Phe116 residue_name_number The interface between the two chains buries 882 Å2 per monomer and is mainly formed by α-helices 2 and 4 and parts of β-sheets 12 and 14, as well as a π–π stacking of the adenine moiety of CoA with Phe116 of the adjacent chain (Fig 4c). RESULTS 69 82 superimposing experimental_method The folds of the two chains in the asymmetric unit are very similar, superimposing with a rmsd of 0.16 Å over 2,306 aligned atom pairs. RESULTS 90 94 rmsd evidence The folds of the two chains in the asymmetric unit are very similar, superimposing with a rmsd of 0.16 Å over 2,306 aligned atom pairs. RESULTS 15 22 helices structure_element The peripheral helices and the short antiparallel β3–4 sheet mediate most of the crystal contacts. RESULTS 31 60 short antiparallel β3–4 sheet structure_element The peripheral helices and the short antiparallel β3–4 sheet mediate most of the crystal contacts. RESULTS 11 22 active site site Details of active site, dimeric assembly, and sequence conservation of PduL. FIG 24 31 dimeric oligomeric_state Details of active site, dimeric assembly, and sequence conservation of PduL. FIG 71 75 PduL protein_type Details of active site, dimeric assembly, and sequence conservation of PduL. FIG 15 26 active site site (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 30 34 PduL protein_type (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 92 100 nitrogen chemical (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 107 113 oxygen chemical (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 119 125 sulfur chemical (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 135 139 zinc chemical (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 189 194 water chemical (a,b) Proposed active site of PduL with relevant residues shown as sticks in atom coloring (nitrogen blue, oxygen red, sulfur yellow), zinc as grey colored spheres and coordinating ordered water molecules in red. FIG 55 65 Coenzyme A chemical Distances between atom centers are indicated in Å. (a) Coenzyme A containing, (b) phosphate-bound structure. FIG 82 97 phosphate-bound protein_state Distances between atom centers are indicated in Å. (a) Coenzyme A containing, (b) phosphate-bound structure. FIG 98 107 structure evidence Distances between atom centers are indicated in Å. (a) Coenzyme A containing, (b) phosphate-bound structure. FIG 16 21 dimer oligomeric_state (c) View of the dimer in the asymmetric unit from the side, domains 1 and 2 colored as in Fig 2 and the two chains differentiated by blue/red versus slate/firebrick. FIG 60 75 domains 1 and 2 structure_element (c) View of the dimer in the asymmetric unit from the side, domains 1 and 2 colored as in Fig 2 and the two chains differentiated by blue/red versus slate/firebrick. FIG 80 84 F116 residue_name_number The asterisk and double arrow marks the location of the π–π interaction between F116 and the CoA base of the other dimer chain. FIG 93 96 CoA chemical The asterisk and double arrow marks the location of the π–π interaction between F116 and the CoA base of the other dimer chain. FIG 115 120 dimer oligomeric_state The asterisk and double arrow marks the location of the π–π interaction between F116 and the CoA base of the other dimer chain. FIG 34 43 structure evidence (d) Surface representation of the structure with indicated conservation (red: high, white: intermediate, yellow: low). FIG 0 29 Size exclusion chromatography experimental_method Size exclusion chromatography of PduL homologs. FIG 33 37 PduL protein_type Size exclusion chromatography of PduL homologs. FIG 9 22 Chromatograms evidence (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 26 31 sPduL protein (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 37 42 rPduL protein (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 52 57 pPduL protein (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 108 110 EP structure_element (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 117 145 nickel affinity purification experimental_method (a)–(c): Chromatograms of sPduL (a), rPduL (b), and pPduL (c) with (orange) or without (blue) the predicted EP, post-nickel affinity purification, applied over a preparative size exclusion column (see Materials and Methods). FIG 9 22 Chromatograms evidence (d)–(f): Chromatograms of sPduL (d), rPduL (e), and pPduL (f) post-preparative size exclusion chromatography with different size fractions separated, applied over an analytical size exclusion column (see Materials and Methods). FIG 26 31 sPduL protein (d)–(f): Chromatograms of sPduL (d), rPduL (e), and pPduL (f) post-preparative size exclusion chromatography with different size fractions separated, applied over an analytical size exclusion column (see Materials and Methods). FIG 37 42 rPduL protein (d)–(f): Chromatograms of sPduL (d), rPduL (e), and pPduL (f) post-preparative size exclusion chromatography with different size fractions separated, applied over an analytical size exclusion column (see Materials and Methods). FIG 52 57 pPduL protein (d)–(f): Chromatograms of sPduL (d), rPduL (e), and pPduL (f) post-preparative size exclusion chromatography with different size fractions separated, applied over an analytical size exclusion column (see Materials and Methods). FIG 79 108 size exclusion chromatography experimental_method (d)–(f): Chromatograms of sPduL (d), rPduL (e), and pPduL (f) post-preparative size exclusion chromatography with different size fractions separated, applied over an analytical size exclusion column (see Materials and Methods). FIG 4 17 chromatograms evidence All chromatograms are cropped to show only the linear range of separation based on standard runs, shown in black squares with a dashed linear trend line. FIG 0 11 Active Site site Active Site Properties RESULTS 0 3 CoA chemical CoA and the metal ions bind between the two domains, presumably in the active site (Figs 2b and 4a). RESULTS 71 82 active site site CoA and the metal ions bind between the two domains, presumably in the active site (Figs 2b and 4a). RESULTS 46 69 X-ray fluorescence scan experimental_method To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 77 85 crystals evidence To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 142 144 Mn chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 146 148 Fe chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 150 152 Co chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 154 156 Ni chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 158 160 Cu chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 166 168 Zn chemical To identify the bound metals, we performed an X-ray fluorescence scan on the crystals at various wavelengths (corresponding to the K-edges of Mn, Fe, Co, Ni, Cu, and Zn). RESULTS 77 81 zinc chemical There was a large signal at the zinc edge, and we tested for the presence of zinc by collecting full data sets before and after the Zn K-edge (1.2861 and 1.2822 Å, respectively). RESULTS 85 162 collecting full data sets before and after the Zn K-edge (1.2861 and 1.2822 Å experimental_method There was a large signal at the zinc edge, and we tested for the presence of zinc by collecting full data sets before and after the Zn K-edge (1.2861 and 1.2822 Å, respectively). RESULTS 76 80 zinc chemical The large differences between the anomalous signals confirm the presence of zinc at both metal sites (S3 Fig). RESULTS 10 14 zinc chemical The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 20 23 Zn1 chemical The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 69 74 His48 residue_name_number The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 76 81 His50 residue_name_number The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 83 89 Glu109 residue_name_number The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 99 102 CoA chemical The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 103 109 sulfur chemical The first zinc ion (Zn1) is in a tetrahedral coordination state with His48, His50, Glu109, and the CoA sulfur (Fig 4a). RESULTS 12 15 Zn2 chemical The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 56 65 conserved protein_state The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 66 75 histidine residue_name The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 86 92 His157 residue_name_number The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 94 100 His159 residue_name_number The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 105 111 His204 residue_name_number The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 130 135 water chemical The second (Zn2) is in octahedral coordination by three conserved histidine residues (His157, His159 and His204) as well as three water molecules (Fig 4a). RESULTS 35 39 zinc chemical The nitrogen atom coordinating the zinc is the Nε in each histidine residue, as is typical for this interaction. RESULTS 58 67 histidine residue_name The nitrogen atom coordinating the zinc is the Nε in each histidine residue, as is typical for this interaction. RESULTS 9 29 crystals were soaked experimental_method When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 35 51 sodium phosphate chemical When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 99 102 CoA chemical When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 120 127 density evidence When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 134 143 phosphate chemical When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 171 182 active site site When the crystals were soaked in a sodium phosphate solution for 2 d prior to data collection, the CoA dissociates, and density for a phosphate molecule is visible at the active site (Table 2, Fig 4b). RESULTS 4 19 phosphate-bound protein_state The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 20 29 structure evidence The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 30 36 aligns experimental_method The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 51 60 CoA-bound protein_state The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 61 70 structure evidence The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 79 83 rmsd evidence The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 109 116 monomer oligomeric_state The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 158 163 dimer oligomeric_state The phosphate-bound structure aligns well with the CoA-bound structure (0.43 Å rmsd over 2,361 atoms for the monomer, 0.83 Å over 5,259 aligned atoms for the dimer). RESULTS 4 13 phosphate chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 28 32 zinc chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 81 84 CoA chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 88 91 Zn1 chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 114 117 Zn2 chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 159 165 waters chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 188 197 phosphate chemical The phosphate contacts both zinc atoms (Fig 4b) and replaces the coordination by CoA at Zn1; the coordination for Zn2 changes from octahedral with three bound waters to tetrahedral with a phosphate ion as one of the ligands (Fig 4b). RESULTS 0 9 Conserved protein_state Conserved Arg103 seems to be involved in maintaining the phosphate in that position. RESULTS 10 16 Arg103 residue_name_number Conserved Arg103 seems to be involved in maintaining the phosphate in that position. RESULTS 57 66 phosphate chemical Conserved Arg103 seems to be involved in maintaining the phosphate in that position. RESULTS 8 12 zinc chemical The two zinc atoms are slightly closer together in the phosphate-bound form (5.8 Å vs 6.3 Å), possibly due to the bridging effect of the phosphate. RESULTS 55 70 phosphate-bound protein_state The two zinc atoms are slightly closer together in the phosphate-bound form (5.8 Å vs 6.3 Å), possibly due to the bridging effect of the phosphate. RESULTS 137 146 phosphate chemical The two zinc atoms are slightly closer together in the phosphate-bound form (5.8 Å vs 6.3 Å), possibly due to the bridging effect of the phosphate. RESULTS 14 23 phosphate chemical An additional phosphate molecule is bound at a crystal contact interface, perhaps accounting for the 14 Å shorter c-axis in the phosphate-bound crystal form (Table 2). RESULTS 128 143 phosphate-bound protein_state An additional phosphate molecule is bound at a crystal contact interface, perhaps accounting for the 14 Å shorter c-axis in the phosphate-bound crystal form (Table 2). RESULTS 21 25 PduL protein_type Oligomeric States of PduL Orthologs Are Influenced by the EP RESULTS 58 60 EP structure_element Oligomeric States of PduL Orthologs Are Influenced by the EP RESULTS 66 71 rPduL protein Interestingly, some of the residues important for dimerization of rPduL, particularly Phe116, are poorly conserved across PduL homologs associated with functionally diverse BMCs (Figs 4c and 3), suggesting that they may have alternative oligomeric states. RESULTS 86 92 Phe116 residue_name_number Interestingly, some of the residues important for dimerization of rPduL, particularly Phe116, are poorly conserved across PduL homologs associated with functionally diverse BMCs (Figs 4c and 3), suggesting that they may have alternative oligomeric states. RESULTS 98 114 poorly conserved protein_state Interestingly, some of the residues important for dimerization of rPduL, particularly Phe116, are poorly conserved across PduL homologs associated with functionally diverse BMCs (Figs 4c and 3), suggesting that they may have alternative oligomeric states. RESULTS 122 126 PduL protein_type Interestingly, some of the residues important for dimerization of rPduL, particularly Phe116, are poorly conserved across PduL homologs associated with functionally diverse BMCs (Figs 4c and 3), suggesting that they may have alternative oligomeric states. RESULTS 173 177 BMCs complex_assembly Interestingly, some of the residues important for dimerization of rPduL, particularly Phe116, are poorly conserved across PduL homologs associated with functionally diverse BMCs (Figs 4c and 3), suggesting that they may have alternative oligomeric states. RESULTS 40 69 size exclusion chromatography experimental_method We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 78 89 full-length protein_state We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 114 121 lacking protein_state We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 126 128 EP structure_element We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 130 133 ΔEP mutant We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 138 143 sPduL protein We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 145 150 rPduL protein We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 156 161 pPduL protein We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 219 223 BMCs complex_assembly We tested this hypothesis by performing size exclusion chromatography on both full-length and truncated variants (lacking the EP, ΔEP) of sPduL, rPduL, and pPduL. These three homologs are found in functionally distinct BMCs (Table 1). RESULTS 30 39 catabolic protein_state It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 40 44 BMCs complex_assembly It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 126 162 aldehyde, and alcohol dehydrogenases protein_type It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 171 174 BMC complex_assembly It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 175 179 PTAC protein_type It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 242 247 shell structure_element It has been proposed that the catabolic BMCs may assemble in a core-first manner, with the luminal enzymes (signature enzyme, aldehyde, and alcohol dehydrogenases and the BMC PTAC) forming an initial bolus, or prometabolosome, around which a shell assembles. RESULTS 73 77 PduL protein_type Given the diversity of signature enzymes (Table 1), it is plausible that PduL orthologs may adopt different oligomeric states that reflect the differences in the proteins being packaged with them in the organelle lumen. RESULTS 170 172 EP structure_element We found that not only did the different orthologs appear to assemble into different oligomeric states, but that quaternary structure was dependent on whether or not the EP was present. RESULTS 0 11 Full-length protein_state Full-length sPduL was unstable in solution—precipitating over time—and eluted throughout the entire volume of a size exclusion column, indicating it was nonspecifically aggregating. RESULTS 12 17 sPduL protein Full-length sPduL was unstable in solution—precipitating over time—and eluted throughout the entire volume of a size exclusion column, indicating it was nonspecifically aggregating. RESULTS 27 29 EP structure_element However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 40 44 1–27 residue_range However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 50 57 removed experimental_method However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 59 68 sPduL ΔEP mutant However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 75 84 truncated protein_state However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 171 178 monomer oligomeric_state However, when the putative EP (residues 1–27) was removed (sPduL ΔEP), the truncated protein was stable and eluted as a single peak (Fig 5a) consistent with the size of a monomer (Fig 5d, blue curve). RESULTS 18 29 full-length protein_state In contrast, both full-length rPduL and pPduL appeared to exist in two distinct oligomeric states (Fig 5b and 5c respectively, orange curves), one form of the approximate size of a dimer and the second, a higher molecular weight oligomer (~150 kDa). RESULTS 30 35 rPduL protein In contrast, both full-length rPduL and pPduL appeared to exist in two distinct oligomeric states (Fig 5b and 5c respectively, orange curves), one form of the approximate size of a dimer and the second, a higher molecular weight oligomer (~150 kDa). RESULTS 40 45 pPduL protein In contrast, both full-length rPduL and pPduL appeared to exist in two distinct oligomeric states (Fig 5b and 5c respectively, orange curves), one form of the approximate size of a dimer and the second, a higher molecular weight oligomer (~150 kDa). RESULTS 181 186 dimer oligomeric_state In contrast, both full-length rPduL and pPduL appeared to exist in two distinct oligomeric states (Fig 5b and 5c respectively, orange curves), one form of the approximate size of a dimer and the second, a higher molecular weight oligomer (~150 kDa). RESULTS 229 237 oligomer oligomeric_state In contrast, both full-length rPduL and pPduL appeared to exist in two distinct oligomeric states (Fig 5b and 5c respectively, orange curves), one form of the approximate size of a dimer and the second, a higher molecular weight oligomer (~150 kDa). RESULTS 5 13 deletion experimental_method Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 30 32 EP structure_element Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 43 47 1–47 residue_range Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 52 57 rPduL protein Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 63 67 1–20 residue_range Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 72 77 pPduL protein Upon deletion of the putative EP (residues 1–47 for rPduL, and 1–20 for pPduL), there was a distinct change in the elution profiles (Fig 5b and 5c respectively, blue curves). RESULTS 0 8 pPduLΔEP mutant pPduLΔEP eluted as two smaller forms, possibly corresponding to a trimer and a monomer. RESULTS 66 72 trimer oligomeric_state pPduLΔEP eluted as two smaller forms, possibly corresponding to a trimer and a monomer. RESULTS 79 86 monomer oligomeric_state pPduLΔEP eluted as two smaller forms, possibly corresponding to a trimer and a monomer. RESULTS 13 21 rPduLΔEP mutant In contrast, rPduLΔEP eluted as one smaller oligomer, possibly a dimer. RESULTS 65 70 dimer oligomeric_state In contrast, rPduLΔEP eluted as one smaller oligomer, possibly a dimer. RESULTS 26 31 rPduL protein We also analyzed purified rPduL and rPduLΔEP by size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) for a complementary approach to assessing oligomeric state. RESULTS 36 44 rPduLΔEP mutant We also analyzed purified rPduL and rPduLΔEP by size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) for a complementary approach to assessing oligomeric state. RESULTS 48 77 size exclusion chromatography experimental_method We also analyzed purified rPduL and rPduLΔEP by size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) for a complementary approach to assessing oligomeric state. RESULTS 91 118 multiangle light scattering experimental_method We also analyzed purified rPduL and rPduLΔEP by size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) for a complementary approach to assessing oligomeric state. RESULTS 120 128 SEC-MALS experimental_method We also analyzed purified rPduL and rPduLΔEP by size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) for a complementary approach to assessing oligomeric state. RESULTS 0 8 SEC-MALS experimental_method SEC-MALS analysis of rPdulΔEP is consistent with a dimer (as observed in the crystal structure) with a weighted average (Mw) and number average (Mn) of the molar mass of 58.4 kDa +/− 11.2% and 58.8 kDa +/− 10.9%, respectively (S4a Fig). RESULTS 21 29 rPdulΔEP mutant SEC-MALS analysis of rPdulΔEP is consistent with a dimer (as observed in the crystal structure) with a weighted average (Mw) and number average (Mn) of the molar mass of 58.4 kDa +/− 11.2% and 58.8 kDa +/− 10.9%, respectively (S4a Fig). RESULTS 51 56 dimer oligomeric_state SEC-MALS analysis of rPdulΔEP is consistent with a dimer (as observed in the crystal structure) with a weighted average (Mw) and number average (Mn) of the molar mass of 58.4 kDa +/− 11.2% and 58.8 kDa +/− 10.9%, respectively (S4a Fig). RESULTS 77 94 crystal structure evidence SEC-MALS analysis of rPdulΔEP is consistent with a dimer (as observed in the crystal structure) with a weighted average (Mw) and number average (Mn) of the molar mass of 58.4 kDa +/− 11.2% and 58.8 kDa +/− 10.9%, respectively (S4a Fig). RESULTS 103 166 weighted average (Mw) and number average (Mn) of the molar mass evidence SEC-MALS analysis of rPdulΔEP is consistent with a dimer (as observed in the crystal structure) with a weighted average (Mw) and number average (Mn) of the molar mass of 58.4 kDa +/− 11.2% and 58.8 kDa +/− 10.9%, respectively (S4a Fig). RESULTS 0 5 rPduL protein rPduL full length runs as Mw = 140.3 kDa +/− 1.2% and Mn = 140.5 kDa +/− 1.2%. RESULTS 6 17 full length protein_state rPduL full length runs as Mw = 140.3 kDa +/− 1.2% and Mn = 140.5 kDa +/− 1.2%. RESULTS 26 28 Mw evidence rPduL full length runs as Mw = 140.3 kDa +/− 1.2% and Mn = 140.5 kDa +/− 1.2%. RESULTS 54 56 Mn evidence rPduL full length runs as Mw = 140.3 kDa +/− 1.2% and Mn = 140.5 kDa +/− 1.2%. RESULTS 43 55 six subunits oligomeric_state This corresponds to an oligomeric state of six subunits (calculated molecular weight of 144 kDa). RESULTS 68 84 molecular weight evidence This corresponds to an oligomeric state of six subunits (calculated molecular weight of 144 kDa). RESULTS 62 64 EP structure_element Collectively, these data strongly suggest that the N-terminal EP of PduL plays a role in defining the quaternary structure of the protein. RESULTS 68 72 PduL protein_type Collectively, these data strongly suggest that the N-terminal EP of PduL plays a role in defining the quaternary structure of the protein. RESULTS 4 7 BMC complex_assembly The BMC shell not only sequesters specific enzymes but also their cofactors, thereby establishing a private cofactor pool dedicated to the encapsulated reactions. DISCUSS 8 13 shell structure_element The BMC shell not only sequesters specific enzymes but also their cofactors, thereby establishing a private cofactor pool dedicated to the encapsulated reactions. DISCUSS 3 12 catabolic protein_state In catabolic BMCs, CoA and NAD+ must be continually recycled within the organelle (Fig 1). DISCUSS 13 17 BMCs complex_assembly In catabolic BMCs, CoA and NAD+ must be continually recycled within the organelle (Fig 1). DISCUSS 19 22 CoA chemical In catabolic BMCs, CoA and NAD+ must be continually recycled within the organelle (Fig 1). DISCUSS 27 31 NAD+ chemical In catabolic BMCs, CoA and NAD+ must be continually recycled within the organelle (Fig 1). DISCUSS 47 69 aldehyde dehydrogenase protein_type Homologs of the predominant cofactor utilizer (aldehyde dehydrogenase) and NAD+ regenerator (alcohol dehydrogenase) have been structurally characterized, but until now structural information was lacking for PduL, which recycles CoA in the organelle lumen. DISCUSS 75 79 NAD+ chemical Homologs of the predominant cofactor utilizer (aldehyde dehydrogenase) and NAD+ regenerator (alcohol dehydrogenase) have been structurally characterized, but until now structural information was lacking for PduL, which recycles CoA in the organelle lumen. DISCUSS 93 114 alcohol dehydrogenase protein_type Homologs of the predominant cofactor utilizer (aldehyde dehydrogenase) and NAD+ regenerator (alcohol dehydrogenase) have been structurally characterized, but until now structural information was lacking for PduL, which recycles CoA in the organelle lumen. DISCUSS 207 211 PduL protein_type Homologs of the predominant cofactor utilizer (aldehyde dehydrogenase) and NAD+ regenerator (alcohol dehydrogenase) have been structurally characterized, but until now structural information was lacking for PduL, which recycles CoA in the organelle lumen. DISCUSS 228 231 CoA chemical Homologs of the predominant cofactor utilizer (aldehyde dehydrogenase) and NAD+ regenerator (alcohol dehydrogenase) have been structurally characterized, but until now structural information was lacking for PduL, which recycles CoA in the organelle lumen. DISCUSS 21 33 housekeeping protein_state Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 34 37 Pta protein_type Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 113 145 ethanolamine-utilizing (EUT) BMC complex_assembly Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 176 180 PduL protein_type Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 224 237 metabolosomes complex_assembly Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 252 261 structure evidence Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 266 277 active site site Curiously, while the housekeeping Pta could provide this function, and indeed does so in the case of one type of ethanolamine-utilizing (EUT) BMC, the evolutionarily unrelated PduL fulfills this function for the majority of metabolosomes using a novel structure and active site for convergent evolution of function. DISCUSS 26 30 PduL protein_type The Tertiary Structure of PduL Is Formed by Discontinuous Segments of Primary Structure DISCUSS 4 13 structure evidence The structure of PduL consists of two β-barrel domains capped by short alpha helical segments (Fig 2b). DISCUSS 17 21 PduL protein_type The structure of PduL consists of two β-barrel domains capped by short alpha helical segments (Fig 2b). DISCUSS 34 54 two β-barrel domains structure_element The structure of PduL consists of two β-barrel domains capped by short alpha helical segments (Fig 2b). DISCUSS 65 93 short alpha helical segments structure_element The structure of PduL consists of two β-barrel domains capped by short alpha helical segments (Fig 2b). DISCUSS 47 60 superimposing experimental_method The two domains are structurally very similar (superimposing with a rmsd of 1.34 Å (over 123 out of 320/348 aligned backbone atoms, S5a Fig). DISCUSS 68 72 rmsd evidence The two domains are structurally very similar (superimposing with a rmsd of 1.34 Å (over 123 out of 320/348 aligned backbone atoms, S5a Fig). DISCUSS 88 98 RHxH motif structure_element However, the amino acid sequences of the two domains are only 16% identical (mainly the RHxH motif, β2 and β10), and 34% similar. DISCUSS 100 102 β2 structure_element However, the amino acid sequences of the two domains are only 16% identical (mainly the RHxH motif, β2 and β10), and 34% similar. DISCUSS 107 110 β10 structure_element However, the amino acid sequences of the two domains are only 16% identical (mainly the RHxH motif, β2 and β10), and 34% similar. DISCUSS 4 13 structure evidence Our structure reveals that the two assigned PF06130 domains (Fig 3) do not form structurally discrete units; this reduces the apparent sequence conservation at the level of primary structure. DISCUSS 44 51 PF06130 structure_element Our structure reveals that the two assigned PF06130 domains (Fig 3) do not form structurally discrete units; this reduces the apparent sequence conservation at the level of primary structure. DISCUSS 4 10 strand structure_element One strand of the domain 1 beta barrel (shown in blue in Fig 2) is contributed by the N-terminus, while the rest of the domain is formed by the residues from the C-terminal half of the protein. DISCUSS 18 26 domain 1 structure_element One strand of the domain 1 beta barrel (shown in blue in Fig 2) is contributed by the N-terminus, while the rest of the domain is formed by the residues from the C-terminal half of the protein. DISCUSS 27 38 beta barrel structure_element One strand of the domain 1 beta barrel (shown in blue in Fig 2) is contributed by the N-terminus, while the rest of the domain is formed by the residues from the C-terminal half of the protein. DISCUSS 162 177 C-terminal half structure_element One strand of the domain 1 beta barrel (shown in blue in Fig 2) is contributed by the N-terminus, while the rest of the domain is formed by the residues from the C-terminal half of the protein. DISCUSS 185 192 protein protein_type One strand of the domain 1 beta barrel (shown in blue in Fig 2) is contributed by the N-terminus, while the rest of the domain is formed by the residues from the C-terminal half of the protein. DISCUSS 5 12 aligned experimental_method When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 16 25 structure evidence When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 31 40 β1 strand structure_element When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 48 60 first domain structure_element When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 102 114 final strand structure_element When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 122 135 second domain structure_element When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 137 139 β9 structure_element When aligned by structure, the β1 strand of the first domain (Fig 2a and 2b, blue) corresponds to the final strand of the second domain (β9), effectively making the domains continuous if the first strand was transplanted to the C-terminus. DISCUSS 39 48 structure evidence Refined domain assignment based on our structure should be able to predict domains of PF06130 homologs much more accurately. DISCUSS 86 93 PF06130 structure_element Refined domain assignment based on our structure should be able to predict domains of PF06130 homologs much more accurately. DISCUSS 38 42 PduL protein_type The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 43 56 barrel domain structure_element The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 102 115 alpha subunit structure_element The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 119 145 ethylbenzene dehydrogenase protein_type The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 156 160 rmsd evidence The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 212 223 beta barrel structure_element The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 232 245 capping helix structure_element The closest structural homolog of the PduL barrel domain is a subdomain of a multienzyme complex, the alpha subunit of ethylbenzene dehydrogenase (S5b Fig, rmsd of 2.26 Å over 226 aligned atoms consisting of one beta barrel and one capping helix). DISCUSS 15 19 PduL protein_type In contrast to PduL, there is only one barrel present in ethylbenzene dehydrogenase, and there is no comparable active site arrangement. DISCUSS 39 45 barrel structure_element In contrast to PduL, there is only one barrel present in ethylbenzene dehydrogenase, and there is no comparable active site arrangement. DISCUSS 57 83 ethylbenzene dehydrogenase protein_type In contrast to PduL, there is only one barrel present in ethylbenzene dehydrogenase, and there is no comparable active site arrangement. DISCUSS 112 123 active site site In contrast to PduL, there is only one barrel present in ethylbenzene dehydrogenase, and there is no comparable active site arrangement. DISCUSS 4 8 PduL protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 42 49 PF06130 structure_element The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 122 126 Acks protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 144 148 PduL protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 166 169 Pta protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 189 210 phosphotransacetylase protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 211 214 Ack protein_type The PduL signature primary structure, two PF06130 domains, occurs in some multidomain proteins, most of them annotated as Acks, suggesting that PduL may also replace Pta in variants of the phosphotransacetylase-Ack pathway. DISCUSS 6 10 PduL protein_type These PduL homologs lack EPs, and their fusion to Ack may have evolved as a way to facilitate substrate channeling between the two enzymes. DISCUSS 20 24 lack protein_state These PduL homologs lack EPs, and their fusion to Ack may have evolved as a way to facilitate substrate channeling between the two enzymes. DISCUSS 25 28 EPs structure_element These PduL homologs lack EPs, and their fusion to Ack may have evolved as a way to facilitate substrate channeling between the two enzymes. DISCUSS 34 39 their protein_type These PduL homologs lack EPs, and their fusion to Ack may have evolved as a way to facilitate substrate channeling between the two enzymes. DISCUSS 50 53 Ack protein_type These PduL homologs lack EPs, and their fusion to Ack may have evolved as a way to facilitate substrate channeling between the two enzymes. DISCUSS 17 29 Metabolosome complex_assembly Implications for Metabolosome Core Assembly DISCUSS 4 7 BMC complex_assembly For BMC-encapsulated proteins to properly function together, they must be targeted to the lumen and assemble into an organization that facilitates substrate/product channeling among the different catalytic sites of the signature and core enzymes. DISCUSS 196 211 catalytic sites site For BMC-encapsulated proteins to properly function together, they must be targeted to the lumen and assemble into an organization that facilitates substrate/product channeling among the different catalytic sites of the signature and core enzymes. DISCUSS 4 24 N-terminal extension structure_element The N-terminal extension on PduL homologs may serve both of these functions. DISCUSS 28 32 PduL protein_type The N-terminal extension on PduL homologs may serve both of these functions. DISCUSS 0 13 The extension structure_element The extension shares many features with previously characterized EPs: it is present only in homologs associated with BMC loci, and it is predicted to form an amphipathic α-helix. DISCUSS 65 68 EPs structure_element The extension shares many features with previously characterized EPs: it is present only in homologs associated with BMC loci, and it is predicted to form an amphipathic α-helix. DISCUSS 117 125 BMC loci gene The extension shares many features with previously characterized EPs: it is present only in homologs associated with BMC loci, and it is predicted to form an amphipathic α-helix. DISCUSS 158 169 amphipathic protein_state The extension shares many features with previously characterized EPs: it is present only in homologs associated with BMC loci, and it is predicted to form an amphipathic α-helix. DISCUSS 170 177 α-helix structure_element The extension shares many features with previously characterized EPs: it is present only in homologs associated with BMC loci, and it is predicted to form an amphipathic α-helix. DISCUSS 14 21 removal experimental_method Moreover, its removal affects the oligomeric state of the protein. DISCUSS 0 2 EP structure_element EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 73 76 BMC complex_assembly EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 99 110 full-length protein_state EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 111 134 propanediol dehydratase protein_type EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 139 165 ethanolamine ammonia-lyase protein_type EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 189 192 PDU complex_assembly EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 197 205 EUT BMCs complex_assembly EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 285 287 EP structure_element EP-mediated oligomerization has been observed for the signature and core BMC enzymes; for example, full-length propanediol dehydratase and ethanolamine ammonia-lyase (signature enzymes for PDU and EUT BMCs) subunits are also insoluble, but become soluble upon removal of the predicted EP. DISCUSS 0 5 sPduL protein sPduL has also previously been reported to localize to inclusion bodies when overexpressed; we show here that this is dependent on the presence of the EP. DISCUSS 77 90 overexpressed experimental_method sPduL has also previously been reported to localize to inclusion bodies when overexpressed; we show here that this is dependent on the presence of the EP. DISCUSS 151 153 EP structure_element sPduL has also previously been reported to localize to inclusion bodies when overexpressed; we show here that this is dependent on the presence of the EP. DISCUSS 23 25 EP structure_element This propensity of the EP to cause proteins to form complexes (Fig 5) might not be a coincidence, but could be a necessary step in the assembly of BMCs. DISCUSS 147 151 BMCs complex_assembly This propensity of the EP to cause proteins to form complexes (Fig 5) might not be a coincidence, but could be a necessary step in the assembly of BMCs. DISCUSS 87 99 metabolosome complex_assembly Structured aggregation of the core enzymes has been proposed to be the initial step in metabolosome assembly and is known to be the first step of β-carboxysome biogenesis, where the core enzyme Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO) is aggregated by the CcmM protein. DISCUSS 194 237 Ribulose Bisphosphate Carboxylase/Oxygenase protein_type Structured aggregation of the core enzymes has been proposed to be the initial step in metabolosome assembly and is known to be the first step of β-carboxysome biogenesis, where the core enzyme Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO) is aggregated by the CcmM protein. DISCUSS 239 246 RuBisCO protein_type Structured aggregation of the core enzymes has been proposed to be the initial step in metabolosome assembly and is known to be the first step of β-carboxysome biogenesis, where the core enzyme Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO) is aggregated by the CcmM protein. DISCUSS 269 273 CcmM protein_type Structured aggregation of the core enzymes has been proposed to be the initial step in metabolosome assembly and is known to be the first step of β-carboxysome biogenesis, where the core enzyme Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO) is aggregated by the CcmM protein. DISCUSS 10 15 CsoS2 protein_type Likewise, CsoS2, a protein in the α-carboxysome core, also aggregates when purified and is proposed to facilitate the nucleation and encapsulation of RuBisCO molecules in the lumen of the organelle. DISCUSS 34 47 α-carboxysome complex_assembly Likewise, CsoS2, a protein in the α-carboxysome core, also aggregates when purified and is proposed to facilitate the nucleation and encapsulation of RuBisCO molecules in the lumen of the organelle. DISCUSS 150 157 RuBisCO protein_type Likewise, CsoS2, a protein in the α-carboxysome core, also aggregates when purified and is proposed to facilitate the nucleation and encapsulation of RuBisCO molecules in the lumen of the organelle. DISCUSS 14 17 EPs structure_element This role for EPs in BMC assembly is in addition to their interaction with shell proteins. DISCUSS 21 24 BMC complex_assembly This role for EPs in BMC assembly is in addition to their interaction with shell proteins. DISCUSS 14 18 PduL protein_type Moreover, the PduL crystal structures offer a clue as to how required cofactors enter the BMC lumen during assembly. DISCUSS 19 37 crystal structures evidence Moreover, the PduL crystal structures offer a clue as to how required cofactors enter the BMC lumen during assembly. DISCUSS 90 93 BMC complex_assembly Moreover, the PduL crystal structures offer a clue as to how required cofactors enter the BMC lumen during assembly. DISCUSS 5 8 CoA chemical Free CoA and NAD+/H could potentially be bound to the enzymes as the core assembles and is encapsulated. DISCUSS 13 17 NAD+ chemical Free CoA and NAD+/H could potentially be bound to the enzymes as the core assembles and is encapsulated. DISCUSS 18 19 H chemical Free CoA and NAD+/H could potentially be bound to the enzymes as the core assembles and is encapsulated. DISCUSS 4 8 PduL protein_type Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 9 17 crystals evidence Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 28 31 CoA chemical Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 59 75 Escherichia coli species Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 123 127 PduL protein_type Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 138 147 CoA-bound protein_state Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 230 238 CoA:PduL complex_assembly Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 250 262 metabolosome complex_assembly Our PduL crystals contained CoA that was captured from the Escherichia coli cytosol, indicating that the “ground state” of PduL is in the CoA-bound form; this could provide an elegantly simple means of guaranteeing a 1:1 ratio of CoA:PduL within the metabolosome lumen. DISCUSS 0 11 Active Site site Active Site Identification and Structural Insights into Catalysis DISCUSS 4 15 active site site The active site of PduL is formed at the interface of the two structural domains (Fig 2b). DISCUSS 19 23 PduL protein_type The active site of PduL is formed at the interface of the two structural domains (Fig 2b). DISCUSS 41 50 interface site The active site of PduL is formed at the interface of the two structural domains (Fig 2b). DISCUSS 73 80 domains structure_element The active site of PduL is formed at the interface of the two structural domains (Fig 2b). DISCUSS 95 106 active site site As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 117 133 highly conserved protein_state As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 164 167 CoA chemical As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 201 206 Ser45 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 208 213 Lys70 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 215 220 Arg97 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 222 227 Leu99 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 229 235 His204 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 237 243 Asn211 residue_name_number As expected, the amino acid sequence conservation is highest in the region around the proposed active site (Fig 4d); highly conserved residues are also involved in CoA binding (Figs 2a and 3, residues Ser45, Lys70, Arg97, Leu99, His204, Asn211). DISCUSS 11 38 metal-coordinating residues site All of the metal-coordinating residues (Fig 2a) are absolutely conserved, implicating them in catalysis or the correct spatial orientation of the substrates. DISCUSS 52 72 absolutely conserved protein_state All of the metal-coordinating residues (Fig 2a) are absolutely conserved, implicating them in catalysis or the correct spatial orientation of the substrates. DISCUSS 0 6 Arg103 residue_name_number Arg103, which contacts the phosphate (Fig 4b), is present in all PduL homologs. DISCUSS 27 36 phosphate chemical Arg103, which contacts the phosphate (Fig 4b), is present in all PduL homologs. DISCUSS 65 69 PduL protein_type Arg103, which contacts the phosphate (Fig 4b), is present in all PduL homologs. DISCUSS 53 56 CoA chemical The close resemblance between the structures binding CoA and phosphate likely indicates that no large changes in protein conformation are involved in catalysis, and that our crystal structures are representative of the active form. DISCUSS 61 70 phosphate chemical The close resemblance between the structures binding CoA and phosphate likely indicates that no large changes in protein conformation are involved in catalysis, and that our crystal structures are representative of the active form. DISCUSS 174 192 crystal structures evidence The close resemblance between the structures binding CoA and phosphate likely indicates that no large changes in protein conformation are involved in catalysis, and that our crystal structures are representative of the active form. DISCUSS 219 225 active protein_state The close resemblance between the structures binding CoA and phosphate likely indicates that no large changes in protein conformation are involved in catalysis, and that our crystal structures are representative of the active form. DISCUSS 49 54 rPduL protein The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 59 64 pPduL protein The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 66 79 propionyl-CoA chemical The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 145 155 nucleotide chemical The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 160 176 pantothenic acid chemical The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 216 229 CoA-thioester chemical The native substrate for the forward reaction of rPduL and pPduL, propionyl-CoA, most likely binds to the enzyme in the same way at the observed nucleotide and pantothenic acid moiety, but the propionyl group in the CoA-thioester might point in a different direction. DISCUSS 11 17 pocket site There is a pocket nearby the active site between the well-conserved residues Ser45 and Ala154, which could accommodate the propionyl group (S6 Fig). DISCUSS 29 40 active site site There is a pocket nearby the active site between the well-conserved residues Ser45 and Ala154, which could accommodate the propionyl group (S6 Fig). DISCUSS 53 67 well-conserved protein_state There is a pocket nearby the active site between the well-conserved residues Ser45 and Ala154, which could accommodate the propionyl group (S6 Fig). DISCUSS 77 82 Ser45 residue_name_number There is a pocket nearby the active site between the well-conserved residues Ser45 and Ala154, which could accommodate the propionyl group (S6 Fig). DISCUSS 87 93 Ala154 residue_name_number There is a pocket nearby the active site between the well-conserved residues Ser45 and Ala154, which could accommodate the propionyl group (S6 Fig). DISCUSS 2 16 homology model experimental_method A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 20 25 sPduL protein A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 69 75 pocket site A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 96 107 active site site A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 140 145 rPduL protein A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 224 237 propionyl-CoA chemical A homology model of sPduL indicates that the residues making up this pocket and the surrounding active site region are identical to that of rPduL, which is not surprising, because these two homologs presumably have the same propionyl-CoA substrate. DISCUSS 4 18 homology model experimental_method The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 22 27 pPduL protein The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 70 76 pocket site The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 127 138 active site site The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 140 145 Gln77 residue_name_number The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 149 154 rPduL protein The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 172 180 tyrosine residue_name The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 182 187 Tyr77 residue_name_number The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 192 197 pPduL protein The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 230 235 pPduL protein The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 263 273 lactyl-CoA chemical The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 331 344 propionyl-CoA chemical The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 365 373 aromatic protein_state The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 374 381 residue structure_element The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 444 448 PduL protein_type The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 465 474 pvm locus gene The homology model of pPduL also has identical residues making up the pocket, but with a key difference in the vicinity of the active site: Gln77 of rPduL is replaced by a tyrosine (Tyr77) in pPduL. The physiological substrate of pPduL (Table 1) is thought to be lactyl-CoA, which contains an additional hydroxyl group relative to propionyl-CoA. The presence of an aromatic residue at this position may underlie the substrate preference of the PduL enzyme from the pvm locus. DISCUSS 27 32 PduLs protein_type Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 44 52 pvm loci gene Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 54 59 Gln77 residue_name_number Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 87 90 Tyr residue_name Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 94 97 Phe residue_name Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 125 128 Gln residue_name Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 132 135 Glu residue_name Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 139 144 PduLs protein_type Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 158 161 BMC complex_assembly Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 181 188 acetyl- chemical Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 192 205 propionyl-CoA chemical Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 209 219 comparison experimental_method Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 227 231 PduL protein_type Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 232 243 active site site Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 282 285 Pta protein_type Indeed, in the majority of PduLs encoded in pvm loci, Gln77 is substituted by either a Tyr or Phe, whereas it is typically a Gln or Glu in PduLs in all other BMC types that degrade acetyl- or propionyl-CoA. A comparison of the PduL active site to that of the functionally identical Pta suggests that the two enzymes have distinctly different mechanisms. DISCUSS 27 30 Pta protein_type The catalytic mechanism of Pta involves the abstraction of a thiol hydrogen by an aspartate residue, resulting in the nucleophilic attack of thiolate upon the carbonyl carbon of acetyl-phosphate, oriented by an arginine and stabilized by a serine —there are no metals involved. DISCUSS 82 91 aspartate residue_name The catalytic mechanism of Pta involves the abstraction of a thiol hydrogen by an aspartate residue, resulting in the nucleophilic attack of thiolate upon the carbonyl carbon of acetyl-phosphate, oriented by an arginine and stabilized by a serine —there are no metals involved. DISCUSS 178 194 acetyl-phosphate chemical The catalytic mechanism of Pta involves the abstraction of a thiol hydrogen by an aspartate residue, resulting in the nucleophilic attack of thiolate upon the carbonyl carbon of acetyl-phosphate, oriented by an arginine and stabilized by a serine —there are no metals involved. DISCUSS 211 219 arginine residue_name The catalytic mechanism of Pta involves the abstraction of a thiol hydrogen by an aspartate residue, resulting in the nucleophilic attack of thiolate upon the carbonyl carbon of acetyl-phosphate, oriented by an arginine and stabilized by a serine —there are no metals involved. DISCUSS 240 246 serine residue_name The catalytic mechanism of Pta involves the abstraction of a thiol hydrogen by an aspartate residue, resulting in the nucleophilic attack of thiolate upon the carbonyl carbon of acetyl-phosphate, oriented by an arginine and stabilized by a serine —there are no metals involved. DISCUSS 20 25 rPduL protein In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 26 35 structure evidence In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 60 69 aspartate residue_name In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 96 107 active site site In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 122 136 well-conserved protein_state In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 137 146 glutamate residue_name In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 162 173 active site site In contrast, in the rPduL structure, there are no conserved aspartate residues in or around the active site, and the only well-conserved glutamate residue in the active site is involved in coordinating one of the metal ions. DISCUSS 44 50 acidic protein_state These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 51 58 residue structure_element These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 100 104 PduL protein_type These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 119 138 dimetal active site site These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 142 146 PduL protein_type These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 212 221 phosphate chemical These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 280 288 acyl-CoA chemical These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 412 426 acyl-phosphate chemical These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 471 483 phosphatases protein_type These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 564 583 dimetal active site site These observations strongly suggest that an acidic residue is not directly involved in catalysis by PduL. Instead, the dimetal active site of PduL may create a nucleophile from one of the hydroxyl groups on free phosphate to attack the carbonyl carbon of the thioester bond of an acyl-CoA. In the reverse direction, the metal ion(s) could stabilize the thiolate anion that would attack the carbonyl carbon of an acyl-phosphate; a similar mechanism has been described for phosphatases where hydroxyl groups or hydroxide ions can act as a base when coordinated by a dimetal active site. DISCUSS 4 14 structures evidence Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 105 109 PduL protein_type Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 111 120 Conserved protein_state Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 137 148 active site site Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 236 242 Ser127 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 244 250 Arg103 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 252 258 Arg194 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 260 266 Gln107 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 268 273 Gln74 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 279 282 Gln residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 283 288 Glu77 residue_name_number Our structures provide the foundation for studies to elucidate the details of the catalytic mechanism of PduL. Conserved residues in the active site that may contribute to substrate binding and/or transition state stabilization include Ser127, Arg103, Arg194, Gln107, Gln74, and Gln/Glu77. DISCUSS 7 22 phosphate-bound protein_state In the phosphate-bound crystal structure, Ser127 and Arg103 appear to position the phosphate (Fig 4b). DISCUSS 23 40 crystal structure evidence In the phosphate-bound crystal structure, Ser127 and Arg103 appear to position the phosphate (Fig 4b). DISCUSS 42 48 Ser127 residue_name_number In the phosphate-bound crystal structure, Ser127 and Arg103 appear to position the phosphate (Fig 4b). DISCUSS 53 59 Arg103 residue_name_number In the phosphate-bound crystal structure, Ser127 and Arg103 appear to position the phosphate (Fig 4b). DISCUSS 83 92 phosphate chemical In the phosphate-bound crystal structure, Ser127 and Arg103 appear to position the phosphate (Fig 4b). DISCUSS 15 21 Arg103 residue_name_number Alternatively, Arg103 might act as a base to render the phosphate more nucleophilic. DISCUSS 56 65 phosphate chemical Alternatively, Arg103 might act as a base to render the phosphate more nucleophilic. DISCUSS 25 30 Gln74 residue_name_number The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 32 35 Gln residue_name_number The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 36 41 Glu77 residue_name_number The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 47 53 Arg194 residue_name_number The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 81 92 active site site The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 101 104 CoA protein_state The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 109 124 phosphate-bound protein_state The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 125 143 crystal structures evidence The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 272 286 acyl-phosphate chemical The functional groups of Gln74, Gln/Glu77, and Arg194 are directed away from the active site in both CoA and phosphate-bound crystal structures and do not appear to be involved in hydrogen bonding with these substrates, although they could be important for positioning an acyl-phosphate. DISCUSS 9 18 CoA-bound protein_state The free CoA-bound form is presumably poised for attack upon an acyl-phosphate, indicating that the enzyme initially binds CoA as opposed to acyl-phosphate. DISCUSS 64 78 acyl-phosphate chemical The free CoA-bound form is presumably poised for attack upon an acyl-phosphate, indicating that the enzyme initially binds CoA as opposed to acyl-phosphate. DISCUSS 123 126 CoA chemical The free CoA-bound form is presumably poised for attack upon an acyl-phosphate, indicating that the enzyme initially binds CoA as opposed to acyl-phosphate. DISCUSS 141 155 acyl-phosphate chemical The free CoA-bound form is presumably poised for attack upon an acyl-phosphate, indicating that the enzyme initially binds CoA as opposed to acyl-phosphate. DISCUSS 53 62 CoA-bound protein_state This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 63 71 crystals evidence This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 100 103 CoA chemical This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 133 138 bound protein_state This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 139 142 CoA chemical This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 152 159 E. coli species This hypothesis is strengthened by the fact that the CoA-bound crystals were obtained without added CoA, indicating that the protein bound CoA from the E. coli expression strain and retained it throughout purification and crystallization. DISCUSS 4 19 phosphate-bound protein_state The phosphate-bound structure indicates that in the opposite reaction direction phosphate is bound first, and then an acyl-CoA enters. DISCUSS 20 29 structure evidence The phosphate-bound structure indicates that in the opposite reaction direction phosphate is bound first, and then an acyl-CoA enters. DISCUSS 80 89 phosphate chemical The phosphate-bound structure indicates that in the opposite reaction direction phosphate is bound first, and then an acyl-CoA enters. DISCUSS 118 126 acyl-CoA chemical The phosphate-bound structure indicates that in the opposite reaction direction phosphate is bound first, and then an acyl-CoA enters. DISCUSS 24 42 crystal structures evidence The two high-resolution crystal structures presented here will serve as the foundation for mechanistic studies on this noncanonical PTAC enzyme to determine how the dimetal active site functions to catalyze both forward and reverse reactions. DISCUSS 132 136 PTAC protein_type The two high-resolution crystal structures presented here will serve as the foundation for mechanistic studies on this noncanonical PTAC enzyme to determine how the dimetal active site functions to catalyze both forward and reverse reactions. DISCUSS 165 184 dimetal active site site The two high-resolution crystal structures presented here will serve as the foundation for mechanistic studies on this noncanonical PTAC enzyme to determine how the dimetal active site functions to catalyze both forward and reverse reactions. DISCUSS 47 51 PduL protein_type Functional, but Not Structural, Convergence of PduL and Pta DISCUSS 56 59 Pta protein_type Functional, but Not Structural, Convergence of PduL and Pta DISCUSS 0 4 PduL protein_type PduL and Pta are mechanistically and structurally distinct enzymes that catalyze the same reaction, a prime example of evolutionary convergence upon a function. DISCUSS 9 12 Pta protein_type PduL and Pta are mechanistically and structurally distinct enzymes that catalyze the same reaction, a prime example of evolutionary convergence upon a function. DISCUSS 155 167 active sites site There are several examples of such functional convergence of enzymes, although typically the enzymes have independently evolved similar, or even identical active sites; for example, the carbonic anhydrase family. DISCUSS 186 204 carbonic anhydrase protein_type There are several examples of such functional convergence of enzymes, although typically the enzymes have independently evolved similar, or even identical active sites; for example, the carbonic anhydrase family. DISCUSS 102 114 active sites site However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 199 202 Pta protein_type However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 207 211 PduL protein_type However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 253 264 β-lactamase protein_type However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 297 308 active site site However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 396 411 metalloproteins protein_type However, apparently less frequent is functional convergence that is supported by distinctly different active sites and accordingly catalytic mechanism, as revealed by comparison of the structures of Pta and PduL. One well-studied example of this is the β-lactamase family of enzymes, in which the active site of Class A and Class C enzymes involve serine-based catalysis, but Class B enzymes are metalloproteins. DISCUSS 27 39 β-lactamases protein_type This is not surprising, as β-lactamases are not so widespread among bacteria and therefore would be expected to have evolved independently several times as a defense mechanism against β-lactam antibiotics. DISCUSS 68 76 bacteria taxonomy_domain This is not surprising, as β-lactamases are not so widespread among bacteria and therefore would be expected to have evolved independently several times as a defense mechanism against β-lactam antibiotics. DISCUSS 20 28 bacteria taxonomy_domain However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 36 39 Pta protein_type However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 81 84 Pta protein_type However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 85 89 PduL protein_type However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 194 211 Pta-encoding gene gene However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 248 252 BMCs complex_assembly However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 295 303 BMC—EUT1 complex_assembly However, nearly all bacteria encode Pta, and it is not immediately clear why the Pta/PduL functional convergence should have evolved: it would seem to be evolutionarily more resourceful for the Pta-encoding gene to be duplicated and repurposed for BMCs, as is apparently the case in one type of BMC—EUT1 (Table 1). DISCUSS 90 94 PduL protein_type There could be some intrinsic biochemical difference between the two enzymes that renders PduL a more attractive candidate for encapsulation in a BMC—for example, PduL might be more amenable to tight packaging, or is better suited for the chemical microenvironment formed within the lumen of the BMC, which can be quite different from the cytosol. DISCUSS 146 149 BMC complex_assembly There could be some intrinsic biochemical difference between the two enzymes that renders PduL a more attractive candidate for encapsulation in a BMC—for example, PduL might be more amenable to tight packaging, or is better suited for the chemical microenvironment formed within the lumen of the BMC, which can be quite different from the cytosol. DISCUSS 163 167 PduL protein_type There could be some intrinsic biochemical difference between the two enzymes that renders PduL a more attractive candidate for encapsulation in a BMC—for example, PduL might be more amenable to tight packaging, or is better suited for the chemical microenvironment formed within the lumen of the BMC, which can be quite different from the cytosol. DISCUSS 296 299 BMC complex_assembly There could be some intrinsic biochemical difference between the two enzymes that renders PduL a more attractive candidate for encapsulation in a BMC—for example, PduL might be more amenable to tight packaging, or is better suited for the chemical microenvironment formed within the lumen of the BMC, which can be quite different from the cytosol. DISCUSS 47 52 PTACs protein_type Further biochemical comparison between the two PTACs will likely yield exciting results that could answer this evolutionary question. DISCUSS 0 4 BMCs complex_assembly BMCs are now known to be widespread among the bacteria and are involved in critical segments of both autotrophic and heterotrophic biochemical pathways that confer to the host organism a competitive (metabolic) advantage in select niches. DISCUSS 46 54 bacteria taxonomy_domain BMCs are now known to be widespread among the bacteria and are involved in critical segments of both autotrophic and heterotrophic biochemical pathways that confer to the host organism a competitive (metabolic) advantage in select niches. DISCUSS 27 39 metabolosome complex_assembly As one of the three common metabolosome core enzymes, the structure of PduL provides a key missing piece to our structural picture of the shared core biochemistry (Fig 1) of functionally diverse catabolic BMCs. DISCUSS 58 67 structure evidence As one of the three common metabolosome core enzymes, the structure of PduL provides a key missing piece to our structural picture of the shared core biochemistry (Fig 1) of functionally diverse catabolic BMCs. DISCUSS 71 75 PduL protein_type As one of the three common metabolosome core enzymes, the structure of PduL provides a key missing piece to our structural picture of the shared core biochemistry (Fig 1) of functionally diverse catabolic BMCs. DISCUSS 195 204 catabolic protein_state As one of the three common metabolosome core enzymes, the structure of PduL provides a key missing piece to our structural picture of the shared core biochemistry (Fig 1) of functionally diverse catabolic BMCs. DISCUSS 205 209 BMCs complex_assembly As one of the three common metabolosome core enzymes, the structure of PduL provides a key missing piece to our structural picture of the shared core biochemistry (Fig 1) of functionally diverse catabolic BMCs. DISCUSS 53 57 PduL protein_type We have observed the oligomeric state differences of PduL to correlate with the presence of an EP, providing new insight into the function of this sequence extension in BMC assembly. DISCUSS 95 97 EP structure_element We have observed the oligomeric state differences of PduL to correlate with the presence of an EP, providing new insight into the function of this sequence extension in BMC assembly. DISCUSS 169 172 BMC complex_assembly We have observed the oligomeric state differences of PduL to correlate with the presence of an EP, providing new insight into the function of this sequence extension in BMC assembly. DISCUSS 42 52 Coenzyme A chemical Moreover, our results suggest a means for Coenzyme A incorporation during metabolosome biogenesis. DISCUSS 74 86 metabolosome complex_assembly Moreover, our results suggest a means for Coenzyme A incorporation during metabolosome biogenesis. DISCUSS 117 121 BMCs complex_assembly A detailed understanding of the underlying principles governing the assembly and internal structural organization of BMCs is a requisite for synthetic biologists to design custom nanoreactors that use BMC architectures as a template. DISCUSS 201 204 BMC complex_assembly A detailed understanding of the underlying principles governing the assembly and internal structural organization of BMCs is a requisite for synthetic biologists to design custom nanoreactors that use BMC architectures as a template. DISCUSS 41 54 metabolosomes complex_assembly Furthermore, given the growing number of metabolosomes implicated in pathogenesis, the PduL structure will be useful in the development of therapeutics. DISCUSS 87 91 PduL protein_type Furthermore, given the growing number of metabolosomes implicated in pathogenesis, the PduL structure will be useful in the development of therapeutics. DISCUSS 92 101 structure evidence Furthermore, given the growing number of metabolosomes implicated in pathogenesis, the PduL structure will be useful in the development of therapeutics. DISCUSS 14 18 PduL protein_type The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS 53 66 metabolosomes complex_assembly The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS 120 124 PduL protein_type The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS 133 136 Pta protein_type The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS 169 172 BMC complex_assembly The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS 249 253 PTAC protein_type The fact that PduL is confined almost exclusively to metabolosomes can be used to develop an inhibitor that blocks only PduL and not Pta as a way to selectively disrupt BMC-based metabolism, while not affecting most commensal organisms that require PTAC activity. DISCUSS