| anno_start anno_end anno_text entity_type sentence section | |
| 0 4 Haem chemical Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance TITLE | |
| 15 27 dimerization oligomeric_state Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance TITLE | |
| 31 37 PGRMC1 protein Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance TITLE | |
| 38 45 Sigma-2 protein Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance TITLE | |
| 0 42 Progesterone-receptor membrane component 1 protein Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 44 50 PGRMC1 protein Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 51 67 Sigma-2 receptor protein Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 74 97 haem-containing protein protein_type Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 118 150 epidermal growth factor receptor protein_type Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 152 156 EGFR protein_type Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 162 178 cytochromes P450 protein_type Progesterone-receptor membrane component 1 (PGRMC1/Sigma-2 receptor) is a haem-containing protein that interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 to regulate cancer proliferation and chemoresistance; its structural basis remains unknown. ABSTRACT | |
| 5 30 crystallographic analyses experimental_method Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 38 44 PGRMC1 protein Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 45 61 cytosolic domain structure_element Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 106 112 stable protein_state Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 113 118 dimer oligomeric_state Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 127 148 stacking interactions bond_interaction Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 167 171 haem chemical Here crystallographic analyses of the PGRMC1 cytosolic domain at 1.95 Å resolution reveal that it forms a stable dimer through stacking interactions of two protruding haem molecules. ABSTRACT | |
| 4 8 haem chemical The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 9 13 iron chemical The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 17 36 five-coordinated by bond_interaction The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 37 43 Tyr113 residue_name_number The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 58 65 surface site The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 73 77 haem chemical The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 87 99 dimerization oligomeric_state The haem iron is five-coordinated by Tyr113, and the open surface of the haem mediates dimerization. ABSTRACT | |
| 0 15 Carbon monoxide chemical Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 17 19 CO chemical Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 37 43 PGRMC1 protein Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 44 56 dimerization oligomeric_state Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 75 98 sixth coordination site site Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 106 110 haem chemical Carbon monoxide (CO) interferes with PGRMC1 dimerization by binding to the sixth coordination site of the haem. ABSTRACT | |
| 0 4 Haem chemical Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 14 20 PGRMC1 protein Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 21 33 dimerization oligomeric_state Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 68 72 EGFR protein_type Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 77 93 cytochromes P450 protein_type Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 201 203 CO chemical Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 207 211 haem chemical Haem-mediated PGRMC1 dimerization is required for interactions with EGFR and cytochromes P450, cancer proliferation and chemoresistance against anti-cancer drugs; these events are attenuated by either CO or haem deprivation in cancer cells. ABSTRACT | |
| 32 44 dimerization oligomeric_state This study demonstrates protein dimerization via haem–haem stacking, which has not been seen in eukaryotes, and provides insights into its functional significance in cancer. ABSTRACT | |
| 49 67 haem–haem stacking bond_interaction This study demonstrates protein dimerization via haem–haem stacking, which has not been seen in eukaryotes, and provides insights into its functional significance in cancer. ABSTRACT | |
| 96 106 eukaryotes taxonomy_domain This study demonstrates protein dimerization via haem–haem stacking, which has not been seen in eukaryotes, and provides insights into its functional significance in cancer. ABSTRACT | |
| 1 7 PGRMC1 protein PGRMC1 binds to EGFR and cytochromes P450, and is known to be involved in cancer proliferation and in drug resistance. ABSTRACT | |
| 17 21 EGFR protein_type PGRMC1 binds to EGFR and cytochromes P450, and is known to be involved in cancer proliferation and in drug resistance. ABSTRACT | |
| 26 42 cytochromes P450 protein_type PGRMC1 binds to EGFR and cytochromes P450, and is known to be involved in cancer proliferation and in drug resistance. ABSTRACT | |
| 32 41 structure evidence Here, the authors determine the structure of the cytosolic domain of PGRMC1, which forms a dimer via haem–haem stacking, and propose how this interaction could be involved in its function. ABSTRACT | |
| 49 65 cytosolic domain structure_element Here, the authors determine the structure of the cytosolic domain of PGRMC1, which forms a dimer via haem–haem stacking, and propose how this interaction could be involved in its function. ABSTRACT | |
| 69 75 PGRMC1 protein Here, the authors determine the structure of the cytosolic domain of PGRMC1, which forms a dimer via haem–haem stacking, and propose how this interaction could be involved in its function. ABSTRACT | |
| 91 96 dimer oligomeric_state Here, the authors determine the structure of the cytosolic domain of PGRMC1, which forms a dimer via haem–haem stacking, and propose how this interaction could be involved in its function. ABSTRACT | |
| 101 119 haem–haem stacking bond_interaction Here, the authors determine the structure of the cytosolic domain of PGRMC1, which forms a dimer via haem–haem stacking, and propose how this interaction could be involved in its function. ABSTRACT | |
| 45 49 haem chemical Much attention has been paid to the roles of haem-iron in cancer development. INTRO | |
| 50 54 iron chemical Much attention has been paid to the roles of haem-iron in cancer development. INTRO | |
| 28 32 haem chemical Increased dietary intake of haem is a risk factor for several types of cancer. INTRO | |
| 29 43 deprivation of protein_state Previous studies showed that deprivation of iron or haem suppresses tumourigenesis. INTRO | |
| 44 48 iron chemical Previous studies showed that deprivation of iron or haem suppresses tumourigenesis. INTRO | |
| 52 56 haem chemical Previous studies showed that deprivation of iron or haem suppresses tumourigenesis. INTRO | |
| 19 34 carbon monoxide chemical On the other hand, carbon monoxide (CO), the gaseous mediator generated by oxidative degradation of haem via haem oxygenase (HO), inhibits tumour growth. INTRO | |
| 36 38 CO chemical On the other hand, carbon monoxide (CO), the gaseous mediator generated by oxidative degradation of haem via haem oxygenase (HO), inhibits tumour growth. INTRO | |
| 100 104 haem chemical On the other hand, carbon monoxide (CO), the gaseous mediator generated by oxidative degradation of haem via haem oxygenase (HO), inhibits tumour growth. INTRO | |
| 109 123 haem oxygenase protein_type On the other hand, carbon monoxide (CO), the gaseous mediator generated by oxidative degradation of haem via haem oxygenase (HO), inhibits tumour growth. INTRO | |
| 125 127 HO protein_type On the other hand, carbon monoxide (CO), the gaseous mediator generated by oxidative degradation of haem via haem oxygenase (HO), inhibits tumour growth. INTRO | |
| 37 41 haem chemical Thus, a tenuous balance between free haem and CO plays key roles in cancer development and chemoresistance, although the underlying mechanisms are not fully understood. INTRO | |
| 46 48 CO chemical Thus, a tenuous balance between free haem and CO plays key roles in cancer development and chemoresistance, although the underlying mechanisms are not fully understood. INTRO | |
| 93 111 affinity nanobeads experimental_method To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 121 125 haem chemical To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 141 183 progesterone-receptor membrane component 1 protein To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 185 191 PGRMC1 protein To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 198 202 haem chemical To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 224 229 mouse taxonomy_domain To gain insight into the underlying mechanisms, we took chemical biological approaches using affinity nanobeads carrying haem and identified progesterone-receptor membrane component 1 (PGRMC1) as a haem-binding protein from mouse liver extracts (Supplementary Fig. 1). INTRO | |
| 0 6 PGRMC1 protein PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 26 67 membrane-associated progesterone receptor protein_type PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 69 73 MAPR protein_type PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 89 107 cytochrome b5-like structure_element PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 108 127 haem-binding region site PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 148 164 highly expressed protein_state PGRMC1 is a member of the membrane-associated progesterone receptor (MAPR) family with a cytochrome b5-like haem-binding region, and is known to be highly expressed in various types of cancers. INTRO | |
| 0 6 PGRMC1 protein PGRMC1 is anchored to the cell membrane through the N-terminal transmembrane helix and interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 (ref). INTRO | |
| 63 82 transmembrane helix structure_element PGRMC1 is anchored to the cell membrane through the N-terminal transmembrane helix and interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 (ref). INTRO | |
| 102 134 epidermal growth factor receptor protein_type PGRMC1 is anchored to the cell membrane through the N-terminal transmembrane helix and interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 (ref). INTRO | |
| 136 140 EGFR protein_type PGRMC1 is anchored to the cell membrane through the N-terminal transmembrane helix and interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 (ref). INTRO | |
| 146 162 cytochromes P450 protein_type PGRMC1 is anchored to the cell membrane through the N-terminal transmembrane helix and interacts with epidermal growth factor receptor (EGFR) and cytochromes P450 (ref). INTRO | |
| 6 12 PGRMC1 protein While PGRMC1 is implicated in cell proliferation and cholesterol biosynthesis, the structural basis on which PGRMC1 exerts its function remains largely unknown. INTRO | |
| 109 115 PGRMC1 protein While PGRMC1 is implicated in cell proliferation and cholesterol biosynthesis, the structural basis on which PGRMC1 exerts its function remains largely unknown. INTRO | |
| 18 24 PGRMC1 protein Here we show that PGRMC1 exhibits a unique haem-dependent dimerization. INTRO | |
| 43 47 haem chemical Here we show that PGRMC1 exhibits a unique haem-dependent dimerization. INTRO | |
| 58 70 dimerization oligomeric_state Here we show that PGRMC1 exhibits a unique haem-dependent dimerization. INTRO | |
| 4 9 dimer oligomeric_state The dimer binds to EGFR and cytochromes P450 to enhance tumour cell proliferation and chemoresistance. INTRO | |
| 19 23 EGFR protein_type The dimer binds to EGFR and cytochromes P450 to enhance tumour cell proliferation and chemoresistance. INTRO | |
| 28 44 cytochromes P450 protein_type The dimer binds to EGFR and cytochromes P450 to enhance tumour cell proliferation and chemoresistance. INTRO | |
| 4 9 dimer oligomeric_state The dimer is dissociated to monomers by physiological levels of CO, suggesting that PGRMC1 serves as a CO-sensitive molecular switch regulating cancer cell proliferation. INTRO | |
| 28 36 monomers oligomeric_state The dimer is dissociated to monomers by physiological levels of CO, suggesting that PGRMC1 serves as a CO-sensitive molecular switch regulating cancer cell proliferation. INTRO | |
| 64 66 CO chemical The dimer is dissociated to monomers by physiological levels of CO, suggesting that PGRMC1 serves as a CO-sensitive molecular switch regulating cancer cell proliferation. INTRO | |
| 84 90 PGRMC1 protein The dimer is dissociated to monomers by physiological levels of CO, suggesting that PGRMC1 serves as a CO-sensitive molecular switch regulating cancer cell proliferation. INTRO | |
| 103 105 CO chemical The dimer is dissociated to monomers by physiological levels of CO, suggesting that PGRMC1 serves as a CO-sensitive molecular switch regulating cancer cell proliferation. INTRO | |
| 0 23 X-ray crystal structure evidence X-ray crystal structure of PGRMC1 RESULTS | |
| 27 33 PGRMC1 protein X-ray crystal structure of PGRMC1 RESULTS | |
| 3 9 solved experimental_method We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 14 31 crystal structure evidence We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 39 49 haem-bound protein_state We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 50 56 PGRMC1 protein We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 57 73 cytosolic domain structure_element We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 79 85 72–195 residue_range We solved the crystal structure of the haem-bound PGRMC1 cytosolic domain (a.a.72–195) at 1.95 Å resolution (Supplementary Fig. 2). RESULTS | |
| 7 18 presence of protein_state In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 19 23 haem chemical In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 25 31 PGRMC1 protein In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 40 47 dimeric oligomeric_state In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 74 98 hydrophobic interactions bond_interaction In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 111 115 haem chemical In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 132 140 monomers oligomeric_state In the presence of haem, PGRMC1 forms a dimeric structure largely through hydrophobic interactions between the haem moieties of two monomers (Fig. 1a, Table 1 and Supplementary Fig. 3; a stereo-structural image is shown in Supplementary Fig 4). RESULTS | |
| 26 32 PGRMC1 protein While the overall fold of PGRMC1 is similar to that of canonical cytochrome b5, their haem irons are coordinated differently. RESULTS | |
| 65 78 cytochrome b5 protein_type While the overall fold of PGRMC1 is similar to that of canonical cytochrome b5, their haem irons are coordinated differently. RESULTS | |
| 86 90 haem chemical While the overall fold of PGRMC1 is similar to that of canonical cytochrome b5, their haem irons are coordinated differently. RESULTS | |
| 3 16 cytochrome b5 protein_type In cytochrome b5, the haem iron is six-coordinated by two axial histidine residues. RESULTS | |
| 22 26 haem chemical In cytochrome b5, the haem iron is six-coordinated by two axial histidine residues. RESULTS | |
| 27 31 iron chemical In cytochrome b5, the haem iron is six-coordinated by two axial histidine residues. RESULTS | |
| 35 53 six-coordinated by bond_interaction In cytochrome b5, the haem iron is six-coordinated by two axial histidine residues. RESULTS | |
| 64 73 histidine residue_name In cytochrome b5, the haem iron is six-coordinated by two axial histidine residues. RESULTS | |
| 6 16 histidines residue_name These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 21 28 missing protein_state These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 32 38 PGRMC1 protein These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 48 52 haem chemical These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 53 57 iron chemical These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 61 80 five-coordinated by bond_interaction These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 81 87 Tyr113 residue_name_number These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 89 93 Y113 residue_name_number These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 95 100 alone protein_state These histidines are missing in PGRMC1, and the haem iron is five-coordinated by Tyr113 (Y113) alone (Fig. 1b and Supplementary Fig. 3). RESULTS | |
| 2 18 homologous helix structure_element A homologous helix that holds haem in cytochrome b5 is longer, shifts away from haem, and does not form a coordinate bond in PGRMC1 (Fig. 1c). RESULTS | |
| 30 34 haem chemical A homologous helix that holds haem in cytochrome b5 is longer, shifts away from haem, and does not form a coordinate bond in PGRMC1 (Fig. 1c). RESULTS | |
| 38 51 cytochrome b5 protein_type A homologous helix that holds haem in cytochrome b5 is longer, shifts away from haem, and does not form a coordinate bond in PGRMC1 (Fig. 1c). RESULTS | |
| 80 84 haem chemical A homologous helix that holds haem in cytochrome b5 is longer, shifts away from haem, and does not form a coordinate bond in PGRMC1 (Fig. 1c). RESULTS | |
| 125 131 PGRMC1 protein A homologous helix that holds haem in cytochrome b5 is longer, shifts away from haem, and does not form a coordinate bond in PGRMC1 (Fig. 1c). RESULTS | |
| 35 39 haem chemical Consequently, the five-coordinated haem of PGRMC1 has an open surface that allows its dimerization through hydrophobic haem–haem stacking. RESULTS | |
| 43 49 PGRMC1 protein Consequently, the five-coordinated haem of PGRMC1 has an open surface that allows its dimerization through hydrophobic haem–haem stacking. RESULTS | |
| 62 69 surface site Consequently, the five-coordinated haem of PGRMC1 has an open surface that allows its dimerization through hydrophobic haem–haem stacking. RESULTS | |
| 86 98 dimerization oligomeric_state Consequently, the five-coordinated haem of PGRMC1 has an open surface that allows its dimerization through hydrophobic haem–haem stacking. RESULTS | |
| 107 137 hydrophobic haem–haem stacking bond_interaction Consequently, the five-coordinated haem of PGRMC1 has an open surface that allows its dimerization through hydrophobic haem–haem stacking. RESULTS | |
| 62 68 Tyr164 residue_name_number Contrary to our finding, Kaluka et al. recently reported that Tyr164 of PGRMC1 is the axial ligand of haem because mutation of this residue impairs haem binding. RESULTS | |
| 72 78 PGRMC1 protein Contrary to our finding, Kaluka et al. recently reported that Tyr164 of PGRMC1 is the axial ligand of haem because mutation of this residue impairs haem binding. RESULTS | |
| 102 106 haem chemical Contrary to our finding, Kaluka et al. recently reported that Tyr164 of PGRMC1 is the axial ligand of haem because mutation of this residue impairs haem binding. RESULTS | |
| 115 123 mutation experimental_method Contrary to our finding, Kaluka et al. recently reported that Tyr164 of PGRMC1 is the axial ligand of haem because mutation of this residue impairs haem binding. RESULTS | |
| 148 152 haem chemical Contrary to our finding, Kaluka et al. recently reported that Tyr164 of PGRMC1 is the axial ligand of haem because mutation of this residue impairs haem binding. RESULTS | |
| 4 19 structural data evidence Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 34 40 Tyr164 residue_name_number Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 74 80 Tyr107 residue_name_number Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 85 91 Lys163 residue_name_number Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 104 119 hydrogen-bonded bond_interaction Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 123 127 haem chemical Our structural data revealed that Tyr164 and a few other residues such as Tyr107 and Lys163 are in fact hydrogen-bonded to haem propionates. RESULTS | |
| 56 63 Tyr 107 residue_name_number This is consistent with observations by Min et al. that Tyr 107 and Tyr113 of PGRMC1 are involved in binding with haem. RESULTS | |
| 68 74 Tyr113 residue_name_number This is consistent with observations by Min et al. that Tyr 107 and Tyr113 of PGRMC1 are involved in binding with haem. RESULTS | |
| 78 84 PGRMC1 protein This is consistent with observations by Min et al. that Tyr 107 and Tyr113 of PGRMC1 are involved in binding with haem. RESULTS | |
| 114 118 haem chemical This is consistent with observations by Min et al. that Tyr 107 and Tyr113 of PGRMC1 are involved in binding with haem. RESULTS | |
| 30 39 conserved protein_state These amino acid residues are conserved among MAPR family members (Supplementary Fig. 5a), suggesting that these proteins share the ability to exhibit haem-dependent dimerization. RESULTS | |
| 46 50 MAPR protein_type These amino acid residues are conserved among MAPR family members (Supplementary Fig. 5a), suggesting that these proteins share the ability to exhibit haem-dependent dimerization. RESULTS | |
| 151 155 haem chemical These amino acid residues are conserved among MAPR family members (Supplementary Fig. 5a), suggesting that these proteins share the ability to exhibit haem-dependent dimerization. RESULTS | |
| 166 178 dimerization oligomeric_state These amino acid residues are conserved among MAPR family members (Supplementary Fig. 5a), suggesting that these proteins share the ability to exhibit haem-dependent dimerization. RESULTS | |
| 0 6 PGRMC1 protein PGRMC1 exhibits haem-dependent dimerization in solution RESULTS | |
| 16 20 haem chemical PGRMC1 exhibits haem-dependent dimerization in solution RESULTS | |
| 31 43 dimerization oligomeric_state PGRMC1 exhibits haem-dependent dimerization in solution RESULTS | |
| 7 13 PGRMC1 protein In the PGRMC1 crystal, two different types of crystal contacts (chain A–A″ and A–B) were observed in addition to the haem-mediated dimer (chain A–A′) (Supplementary Figs 3 and 6a). RESULTS | |
| 14 21 crystal evidence In the PGRMC1 crystal, two different types of crystal contacts (chain A–A″ and A–B) were observed in addition to the haem-mediated dimer (chain A–A′) (Supplementary Figs 3 and 6a). RESULTS | |
| 117 121 haem chemical In the PGRMC1 crystal, two different types of crystal contacts (chain A–A″ and A–B) were observed in addition to the haem-mediated dimer (chain A–A′) (Supplementary Figs 3 and 6a). RESULTS | |
| 131 136 dimer oligomeric_state In the PGRMC1 crystal, two different types of crystal contacts (chain A–A″ and A–B) were observed in addition to the haem-mediated dimer (chain A–A′) (Supplementary Figs 3 and 6a). RESULTS | |
| 16 20 haem chemical To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 30 42 dimerization oligomeric_state To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 46 52 PGRMC1 protein To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 89 98 structure evidence To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 102 105 apo protein_state To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 111 121 haem-bound protein_state To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 122 128 PGMRC1 protein To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 132 174 two-dimensional nuclear magnetic resonance experimental_method To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 176 179 NMR experimental_method To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 187 274 heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy experimental_method To confirm that haem-assisted dimerization of PGRMC1 occurs in solution, we analysed the structure of apo- and haem-bound PGMRC1 by two-dimensional nuclear magnetic resonance (NMR) using heteronuclear single-quantum coherence and transverse relaxation-optimized spectroscopy (Supplementary Figs 6b and 7). RESULTS | |
| 0 3 NMR experimental_method NMR signals from some amino acid residues of PGRMC1 disappeared due to the paramagnetic relaxation effect of haem (Supplementary Figs 6b); these residues were located in the haem-binding region. RESULTS | |
| 45 51 PGRMC1 protein NMR signals from some amino acid residues of PGRMC1 disappeared due to the paramagnetic relaxation effect of haem (Supplementary Figs 6b); these residues were located in the haem-binding region. RESULTS | |
| 109 113 haem chemical NMR signals from some amino acid residues of PGRMC1 disappeared due to the paramagnetic relaxation effect of haem (Supplementary Figs 6b); these residues were located in the haem-binding region. RESULTS | |
| 174 193 haem-binding region site NMR signals from some amino acid residues of PGRMC1 disappeared due to the paramagnetic relaxation effect of haem (Supplementary Figs 6b); these residues were located in the haem-binding region. RESULTS | |
| 5 20 chemical shifts evidence When chemical shifts of apo- and haem-bound forms of PGMRC1 were compared, some amino acid residues close to those which disappeared because of the paramagnetic relaxation effect of haem exhibit notable chemical shifts (Supplementary Fig. 6a,b; dark yellow). RESULTS | |
| 24 27 apo protein_state When chemical shifts of apo- and haem-bound forms of PGMRC1 were compared, some amino acid residues close to those which disappeared because of the paramagnetic relaxation effect of haem exhibit notable chemical shifts (Supplementary Fig. 6a,b; dark yellow). RESULTS | |
| 33 43 haem-bound protein_state When chemical shifts of apo- and haem-bound forms of PGMRC1 were compared, some amino acid residues close to those which disappeared because of the paramagnetic relaxation effect of haem exhibit notable chemical shifts (Supplementary Fig. 6a,b; dark yellow). RESULTS | |
| 53 59 PGMRC1 protein When chemical shifts of apo- and haem-bound forms of PGMRC1 were compared, some amino acid residues close to those which disappeared because of the paramagnetic relaxation effect of haem exhibit notable chemical shifts (Supplementary Fig. 6a,b; dark yellow). RESULTS | |
| 182 186 haem chemical When chemical shifts of apo- and haem-bound forms of PGMRC1 were compared, some amino acid residues close to those which disappeared because of the paramagnetic relaxation effect of haem exhibit notable chemical shifts (Supplementary Fig. 6a,b; dark yellow). RESULTS | |
| 16 26 interfaces site However, at the interfaces of the other possible dimeric structures (Supplementary Fig. 6a, chain A–A″; cyan and chain A–B; violet), no significant difference was observed. RESULTS | |
| 49 56 dimeric oligomeric_state However, at the interfaces of the other possible dimeric structures (Supplementary Fig. 6a, chain A–A″; cyan and chain A–B; violet), no significant difference was observed. RESULTS | |
| 57 67 structures evidence However, at the interfaces of the other possible dimeric structures (Supplementary Fig. 6a, chain A–A″; cyan and chain A–B; violet), no significant difference was observed. RESULTS | |
| 13 40 free energy of dissociation evidence Furthermore, free energy of dissociation predicted by PISA suggested that the haem-mediated dimer is stable in solution while the other potential interactions are not. RESULTS | |
| 54 58 PISA experimental_method Furthermore, free energy of dissociation predicted by PISA suggested that the haem-mediated dimer is stable in solution while the other potential interactions are not. RESULTS | |
| 78 82 haem chemical Furthermore, free energy of dissociation predicted by PISA suggested that the haem-mediated dimer is stable in solution while the other potential interactions are not. RESULTS | |
| 92 97 dimer oligomeric_state Furthermore, free energy of dissociation predicted by PISA suggested that the haem-mediated dimer is stable in solution while the other potential interactions are not. RESULTS | |
| 101 107 stable protein_state Furthermore, free energy of dissociation predicted by PISA suggested that the haem-mediated dimer is stable in solution while the other potential interactions are not. RESULTS | |
| 56 62 PGRMC1 protein We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 71 74 NMR experimental_method We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 100 114 TALOS+ program experimental_method We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 223 226 apo protein_state We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 232 242 haem-bound protein_state We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 252 258 PGRMC1 protein We also attempted to predict the secondary structure of PGRMC1 through NMR data by calculating with TALOS+ program (Supplementary Fig. 8); the prediction suggested that the overall secondary structure is comparable between apo- and haem-bound forms of PGRMC1 in solution. RESULTS | |
| 16 20 haem chemical We analysed the haem-dependent dimerization of the PGRMC1 cytosolic domain (a.a.44–195) in solution (Fig. 2 and Table 2). RESULTS | |
| 31 43 dimerization oligomeric_state We analysed the haem-dependent dimerization of the PGRMC1 cytosolic domain (a.a.44–195) in solution (Fig. 2 and Table 2). RESULTS | |
| 51 57 PGRMC1 protein We analysed the haem-dependent dimerization of the PGRMC1 cytosolic domain (a.a.44–195) in solution (Fig. 2 and Table 2). RESULTS | |
| 58 74 cytosolic domain structure_element We analysed the haem-dependent dimerization of the PGRMC1 cytosolic domain (a.a.44–195) in solution (Fig. 2 and Table 2). RESULTS | |
| 80 86 44–195 residue_range We analysed the haem-dependent dimerization of the PGRMC1 cytosolic domain (a.a.44–195) in solution (Fig. 2 and Table 2). RESULTS | |
| 0 17 Mass spectrometry experimental_method Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 19 21 MS experimental_method Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 38 62 non-denaturing condition experimental_method Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 85 88 apo protein_state Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 89 96 monomer oligomeric_state Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 97 103 PGRMC1 protein Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 116 128 dimerization oligomeric_state Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 145 149 haem chemical Mass spectrometry (MS) analyses under non-denaturing condition demonstrated that the apo-monomer PGRMC1 resulted in dimerization by binding with haem (Fig. 2a). RESULTS | |
| 26 40 disulfide bond ptm It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 53 59 Cys129 residue_name_number It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 88 95 crystal evidence It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 99 105 PGRMC1 protein It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 123 129 Cys129 residue_name_number It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 133 146 not conserved protein_state It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 157 161 MAPR protein_type It should be noted that a disulfide bond between two Cys129 residues is observed in the crystal of PGRMC1 (Fig. 1a), while Cys129 is not conserved among the MAPR family proteins (Supplementary Fig. 5a). RESULTS | |
| 54 68 disulfide bond ptm This observation led us to examine whether or not the disulfide bond contributes to PGRMC1 dimerization. RESULTS | |
| 84 90 PGRMC1 protein This observation led us to examine whether or not the disulfide bond contributes to PGRMC1 dimerization. RESULTS | |
| 91 103 dimerization oligomeric_state This observation led us to examine whether or not the disulfide bond contributes to PGRMC1 dimerization. RESULTS | |
| 0 2 MS experimental_method MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 18 43 non-denaturing conditions experimental_method MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 68 77 Cys129Ser mutant MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 79 84 C129S mutant MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 86 92 mutant protein_state MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 96 105 dimerized protein_state MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 113 124 presence of protein_state MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 125 129 haem chemical MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 151 155 haem chemical MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 165 177 dimerization oligomeric_state MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 181 187 PGRMC1 protein MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 216 230 disulfide bond ptm MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 245 251 Cys129 residue_name_number MS analyses under non-denaturing conditions clearly showed that the Cys129Ser (C129S) mutant is dimerized in the presence of haem, indicating that the haem-mediated dimerization of PGRMC1 occurs independently of the disulfide bond formation via Cys129 (Fig. 2a). RESULTS | |
| 17 19 MS experimental_method Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 35 56 denaturing conditions experimental_method Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 69 73 haem chemical Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 83 89 PGRMC1 protein Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 90 95 dimer oligomeric_state Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 127 134 monomer oligomeric_state Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 152 164 dimerization oligomeric_state Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 223 237 disulfide bond ptm Supporting this, MS analyses under denaturing conditions showed that haem-mediated PGRMC1 dimer is completely dissociated into monomer, indicating that dimerization of this kind is not mediated by any covalent bond such as disulfide bond (Supplementary Fig. 9). RESULTS | |
| 21 25 haem chemical We also analysed the haem-dependent dimerization of PGRMC1 by diffusion-ordered NMR spectroscopy (DOSY) analyses (Table 2, Supplementary Fig. 10). RESULTS | |
| 36 48 dimerization oligomeric_state We also analysed the haem-dependent dimerization of PGRMC1 by diffusion-ordered NMR spectroscopy (DOSY) analyses (Table 2, Supplementary Fig. 10). RESULTS | |
| 52 58 PGRMC1 protein We also analysed the haem-dependent dimerization of PGRMC1 by diffusion-ordered NMR spectroscopy (DOSY) analyses (Table 2, Supplementary Fig. 10). RESULTS | |
| 62 96 diffusion-ordered NMR spectroscopy experimental_method We also analysed the haem-dependent dimerization of PGRMC1 by diffusion-ordered NMR spectroscopy (DOSY) analyses (Table 2, Supplementary Fig. 10). RESULTS | |
| 98 102 DOSY experimental_method We also analysed the haem-dependent dimerization of PGRMC1 by diffusion-ordered NMR spectroscopy (DOSY) analyses (Table 2, Supplementary Fig. 10). RESULTS | |
| 31 50 hydrodynamic radius evidence The results suggested that the hydrodynamic radius of haem-bound PGRMC1 is larger than that of apo-PGRMC1. RESULTS | |
| 54 64 haem-bound protein_state The results suggested that the hydrodynamic radius of haem-bound PGRMC1 is larger than that of apo-PGRMC1. RESULTS | |
| 65 71 PGRMC1 protein The results suggested that the hydrodynamic radius of haem-bound PGRMC1 is larger than that of apo-PGRMC1. RESULTS | |
| 95 98 apo protein_state The results suggested that the hydrodynamic radius of haem-bound PGRMC1 is larger than that of apo-PGRMC1. RESULTS | |
| 99 105 PGRMC1 protein The results suggested that the hydrodynamic radius of haem-bound PGRMC1 is larger than that of apo-PGRMC1. RESULTS | |
| 52 64 dimerization oligomeric_state To further evaluate changes in molecular weights in dimerization of PGRMC1, sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis was carried out. RESULTS | |
| 68 74 PGRMC1 protein To further evaluate changes in molecular weights in dimerization of PGRMC1, sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis was carried out. RESULTS | |
| 76 129 sedimentation velocity analytical ultracentrifugation experimental_method To further evaluate changes in molecular weights in dimerization of PGRMC1, sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis was carried out. RESULTS | |
| 131 137 SV-AUC experimental_method To further evaluate changes in molecular weights in dimerization of PGRMC1, sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis was carried out. RESULTS | |
| 12 21 wild-type protein_state Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 23 25 wt protein_state Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 27 30 apo protein_state Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 31 37 PGRMC1 protein Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 66 73 monomer oligomeric_state Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 79 83 haem chemical Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 92 98 PGRMC1 protein Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 118 123 dimer oligomeric_state Whereas the wild-type (wt) apo-PGRMC1 appeared at a 1.9 S peak as monomer, the haem-binding PGRMC1 was converted into dimer at a 3.1 S peak (Fig. 2b). RESULTS | |
| 15 20 C129S mutant Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 21 27 mutant protein_state Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 31 37 PGRMC1 protein Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 53 60 monomer oligomeric_state Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 64 69 dimer oligomeric_state Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 84 88 haem chemical Similarly, the C129S mutant of PGRMC1 converted from monomer to dimer by binding to haem (Fig. 2b). RESULTS | |
| 0 6 SV-AUC experimental_method SV-AUC analyses also allowed us to examine the stability of haem/PGRMC1 dimer. RESULTS | |
| 60 64 haem chemical SV-AUC analyses also allowed us to examine the stability of haem/PGRMC1 dimer. RESULTS | |
| 65 71 PGRMC1 protein SV-AUC analyses also allowed us to examine the stability of haem/PGRMC1 dimer. RESULTS | |
| 72 77 dimer oligomeric_state SV-AUC analyses also allowed us to examine the stability of haem/PGRMC1 dimer. RESULTS | |
| 68 78 haem-bound protein_state To this end, we used different concentrations (3.5–147 μmol l−1) of haem-bound PGRMC1 protein (a.a. 72–195), which were identical to that used in the crystallographic analysis. RESULTS | |
| 79 85 PGRMC1 protein To this end, we used different concentrations (3.5–147 μmol l−1) of haem-bound PGRMC1 protein (a.a. 72–195), which were identical to that used in the crystallographic analysis. RESULTS | |
| 100 106 72–195 residue_range To this end, we used different concentrations (3.5–147 μmol l−1) of haem-bound PGRMC1 protein (a.a. 72–195), which were identical to that used in the crystallographic analysis. RESULTS | |
| 150 175 crystallographic analysis experimental_method To this end, we used different concentrations (3.5–147 μmol l−1) of haem-bound PGRMC1 protein (a.a. 72–195), which were identical to that used in the crystallographic analysis. RESULTS | |
| 4 30 sedimentation coefficients evidence The sedimentation coefficients calculated on the basis of the crystal structure were 1.71 S for monomer and 2.56 S for dimer (Supplementary Fig. 11, upper panel). RESULTS | |
| 62 79 crystal structure evidence The sedimentation coefficients calculated on the basis of the crystal structure were 1.71 S for monomer and 2.56 S for dimer (Supplementary Fig. 11, upper panel). RESULTS | |
| 96 103 monomer oligomeric_state The sedimentation coefficients calculated on the basis of the crystal structure were 1.71 S for monomer and 2.56 S for dimer (Supplementary Fig. 11, upper panel). RESULTS | |
| 119 124 dimer oligomeric_state The sedimentation coefficients calculated on the basis of the crystal structure were 1.71 S for monomer and 2.56 S for dimer (Supplementary Fig. 11, upper panel). RESULTS | |
| 28 34 PGRMC1 protein The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 35 40 dimer oligomeric_state The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 65 72 monomer oligomeric_state The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 162 164 Kd evidence The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 174 178 haem chemical The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 188 193 dimer oligomeric_state The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 197 203 PGRMC1 protein The results showed that the PGRMC1 dimer is not dissociated into monomer at all concentrations examined (Supplementary Fig. 11, lower panel), suggesting that the Kd value of haem-mediated dimer of PGRMC1 is under 3.5 μmol l−1. RESULTS | |
| 38 44 PGRMC1 protein A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 45 50 dimer oligomeric_state A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 77 83 dimers oligomeric_state A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 87 107 extracellular domain structure_element A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 111 128 membrane proteins protein_type A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 137 157 Toll like receptor 9 protein A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 159 171 dimerization oligomeric_state A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 172 174 Kd evidence A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 202 220 plexin A2 receptor protein A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 222 234 dimerization oligomeric_state A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 235 237 Kd evidence A value of this kind implies that the PGRMC1 dimer is more stable than other dimers of extracellular domain of membrane proteins such as Toll like receptor 9 (dimerization Kd of 20 μmol l−1) (ref.) and plexin A2 receptor (dimerization Kd higher than 300 μmol l−1) (ref.). RESULTS | |
| 43 46 apo protein_state The current analytical data confirmed that apo-PGRMC1 monomer converts into dimer by binding to haem in solution (Table 2). RESULTS | |
| 47 53 PGRMC1 protein The current analytical data confirmed that apo-PGRMC1 monomer converts into dimer by binding to haem in solution (Table 2). RESULTS | |
| 54 61 monomer oligomeric_state The current analytical data confirmed that apo-PGRMC1 monomer converts into dimer by binding to haem in solution (Table 2). RESULTS | |
| 76 81 dimer oligomeric_state The current analytical data confirmed that apo-PGRMC1 monomer converts into dimer by binding to haem in solution (Table 2). RESULTS | |
| 96 100 haem chemical The current analytical data confirmed that apo-PGRMC1 monomer converts into dimer by binding to haem in solution (Table 2). RESULTS | |
| 18 44 haem titration experiments experimental_method We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 50 54 haem chemical We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 66 72 PGRMC1 protein We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 100 102 Kd evidence We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 157 182 iron regulatory protein 2 protein We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 242 246 haem chemical We also showed by haem titration experiments that haem binding to PGRMC1 was of low affinity with a Kd value of 50 nmol l−1; this is comparable with that of iron regulatory protein 2, which is known to be regulated by intracellular levels of haem (Fig. 2c and Supplementary Table 1). RESULTS | |
| 58 64 PGRMC1 protein These results raised the possibility that the function of PGRMC1 is regulated by intracellular haem concentrations. RESULTS | |
| 95 99 haem chemical These results raised the possibility that the function of PGRMC1 is regulated by intracellular haem concentrations. RESULTS | |
| 0 2 CO chemical CO inhibits haem-dependent dimerization of PGRMC1 RESULTS | |
| 12 16 haem chemical CO inhibits haem-dependent dimerization of PGRMC1 RESULTS | |
| 27 39 dimerization oligomeric_state CO inhibits haem-dependent dimerization of PGRMC1 RESULTS | |
| 43 49 PGRMC1 protein CO inhibits haem-dependent dimerization of PGRMC1 RESULTS | |
| 0 25 Crystallographic analyses experimental_method Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 40 46 Tyr113 residue_name_number Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 50 56 PGRMC1 protein Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 80 84 haem chemical Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 104 108 haem chemical Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 119 131 dimerization oligomeric_state Crystallographic analyses revealed that Tyr113 of PGRMC1 is an axial ligand for haem and contributes to haem-dependent dimerization (Fig. 1a). RESULTS | |
| 12 30 UV-visible spectra evidence Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 49 53 heme chemical Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 57 63 PGRMC1 protein Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 82 88 ferric protein_state Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 92 99 ferrous protein_state Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 121 123 CO chemical Analysis of UV-visible spectra revealed that the heme of PGRMC1 is reducible from ferric to ferrous state, thus allowing CO binding (Fig. 3a). RESULTS | |
| 17 36 UV-visible spectrum evidence Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 44 53 wild type protein_state Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 54 60 PGRMC1 protein Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 89 94 C129S mutant Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 95 101 mutant protein_state Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 105 111 PGRMC1 protein Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 121 130 R/Z ratio evidence Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 250 267 fully loaded with protein_state Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 268 272 haem chemical Furthermore, the UV-visible spectrum of the wild type PGRMC1 was the same as that of the C129S mutant of PGRMC1, and the R/Z ratio determined by the intensities between the Soret band (394 nm) peak and the 274-nm peak showed that these proteins were fully loaded with haem (Supplementary Fig. 12). RESULTS | |
| 16 22 ferric protein_state Analysis of the ferric form of PGRMC1 using resonance Raman spectroscopy (Supplementary Fig. 13) showed that the relative intensity of oxidation and spin state marker bands (ν4 and ν3) is close to 1.0, which is consistent with it being a haem protein with a proximal Tyr coordination. RESULTS | |
| 31 37 PGRMC1 protein Analysis of the ferric form of PGRMC1 using resonance Raman spectroscopy (Supplementary Fig. 13) showed that the relative intensity of oxidation and spin state marker bands (ν4 and ν3) is close to 1.0, which is consistent with it being a haem protein with a proximal Tyr coordination. RESULTS | |
| 44 72 resonance Raman spectroscopy experimental_method Analysis of the ferric form of PGRMC1 using resonance Raman spectroscopy (Supplementary Fig. 13) showed that the relative intensity of oxidation and spin state marker bands (ν4 and ν3) is close to 1.0, which is consistent with it being a haem protein with a proximal Tyr coordination. RESULTS | |
| 238 242 haem chemical Analysis of the ferric form of PGRMC1 using resonance Raman spectroscopy (Supplementary Fig. 13) showed that the relative intensity of oxidation and spin state marker bands (ν4 and ν3) is close to 1.0, which is consistent with it being a haem protein with a proximal Tyr coordination. RESULTS | |
| 267 270 Tyr residue_name Analysis of the ferric form of PGRMC1 using resonance Raman spectroscopy (Supplementary Fig. 13) showed that the relative intensity of oxidation and spin state marker bands (ν4 and ν3) is close to 1.0, which is consistent with it being a haem protein with a proximal Tyr coordination. RESULTS | |
| 11 22 Raman shift evidence A specific Raman shift peaking at vFe–CO=500 cm−1 demonstrated that the CO-bound haem of PGRMC1 is six-coordinated (Supplementary Fig. 13). RESULTS | |
| 72 80 CO-bound protein_state A specific Raman shift peaking at vFe–CO=500 cm−1 demonstrated that the CO-bound haem of PGRMC1 is six-coordinated (Supplementary Fig. 13). RESULTS | |
| 81 85 haem chemical A specific Raman shift peaking at vFe–CO=500 cm−1 demonstrated that the CO-bound haem of PGRMC1 is six-coordinated (Supplementary Fig. 13). RESULTS | |
| 89 95 PGRMC1 protein A specific Raman shift peaking at vFe–CO=500 cm−1 demonstrated that the CO-bound haem of PGRMC1 is six-coordinated (Supplementary Fig. 13). RESULTS | |
| 6 12 PGRMC1 protein Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 13 25 dimerization oligomeric_state Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 44 51 surface site Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 55 59 haem chemical Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 94 100 Tyr113 residue_name_number Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 115 117 CO chemical Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 146 153 dimeric oligomeric_state Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 154 163 structure evidence Since PGRMC1 dimerization involves the open surface of haem on the opposite side of the axial Tyr113, no space for CO binding is available in the dimeric structure (Fig. 3b). RESULTS | |
| 27 29 CO chemical This prompted us to ask if CO binding to haem causes dissociation of the PGRMC1 dimer. RESULTS | |
| 41 45 haem chemical This prompted us to ask if CO binding to haem causes dissociation of the PGRMC1 dimer. RESULTS | |
| 73 79 PGRMC1 protein This prompted us to ask if CO binding to haem causes dissociation of the PGRMC1 dimer. RESULTS | |
| 80 85 dimer oligomeric_state This prompted us to ask if CO binding to haem causes dissociation of the PGRMC1 dimer. RESULTS | |
| 12 41 gel filtration chromatography experimental_method Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 92 101 wild-type protein_state Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 110 115 C129S mutant Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 116 122 mutant protein_state Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 126 132 PGRMC1 protein Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 157 161 haem chemical Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 165 168 apo protein_state Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 169 175 PGRMC1 protein Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 217 221 iron chemical Analysis by gel filtration chromatography revealed that the relative molecular sizes of the wild-type and the C129S mutant of PGRMC1 are increased by adding haem to apo-PGRMC1 regardless of the oxidation state of the iron (Fig. 3c), which is in agreement with the results in Table 1. RESULTS | |
| 0 2 CO chemical CO application to ferrous PGRMC1 abolished the haem-dependent increase in its molecular size. RESULTS | |
| 18 25 ferrous protein_state CO application to ferrous PGRMC1 abolished the haem-dependent increase in its molecular size. RESULTS | |
| 26 32 PGRMC1 protein CO application to ferrous PGRMC1 abolished the haem-dependent increase in its molecular size. RESULTS | |
| 47 51 haem chemical CO application to ferrous PGRMC1 abolished the haem-dependent increase in its molecular size. RESULTS | |
| 37 48 presence of protein_state Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 49 59 dithionite chemical Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 73 91 UV-visible spectra evidence Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 107 109 CO chemical Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 123 134 haem-PGRMC1 complex_assembly Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 138 144 stable protein_state Under this reducing condition in the presence of dithionite, analyses of UV-visible spectra indicated that CO-binding with haem-PGRMC1 is stable, showing only 20% reduction of the absorbance at 412 nm within 2 h (Supplementary Fig. 14). RESULTS | |
| 17 26 Tyr113Phe mutant Furthermore, the Tyr113Phe (Y113F) mutant of PGRMC1 was not responsive to haem. RESULTS | |
| 28 33 Y113F mutant Furthermore, the Tyr113Phe (Y113F) mutant of PGRMC1 was not responsive to haem. RESULTS | |
| 35 41 mutant protein_state Furthermore, the Tyr113Phe (Y113F) mutant of PGRMC1 was not responsive to haem. RESULTS | |
| 45 51 PGRMC1 protein Furthermore, the Tyr113Phe (Y113F) mutant of PGRMC1 was not responsive to haem. RESULTS | |
| 74 78 haem chemical Furthermore, the Tyr113Phe (Y113F) mutant of PGRMC1 was not responsive to haem. RESULTS | |
| 27 29 CO chemical These results suggest that CO favours the six-coordinate form of haem and interferes with the haem-mediated dimerization of PGRMC1. RESULTS | |
| 65 69 haem chemical These results suggest that CO favours the six-coordinate form of haem and interferes with the haem-mediated dimerization of PGRMC1. RESULTS | |
| 94 98 haem chemical These results suggest that CO favours the six-coordinate form of haem and interferes with the haem-mediated dimerization of PGRMC1. RESULTS | |
| 108 120 dimerization oligomeric_state These results suggest that CO favours the six-coordinate form of haem and interferes with the haem-mediated dimerization of PGRMC1. RESULTS | |
| 124 130 PGRMC1 protein These results suggest that CO favours the six-coordinate form of haem and interferes with the haem-mediated dimerization of PGRMC1. RESULTS | |
| 37 39 CO chemical To examine the inhibitory effects of CO on haem-mediated PGRMC1 dimerization, SV-AUC analysis was carried out. RESULTS | |
| 43 47 haem chemical To examine the inhibitory effects of CO on haem-mediated PGRMC1 dimerization, SV-AUC analysis was carried out. RESULTS | |
| 57 63 PGRMC1 protein To examine the inhibitory effects of CO on haem-mediated PGRMC1 dimerization, SV-AUC analysis was carried out. RESULTS | |
| 64 76 dimerization oligomeric_state To examine the inhibitory effects of CO on haem-mediated PGRMC1 dimerization, SV-AUC analysis was carried out. RESULTS | |
| 78 84 SV-AUC experimental_method To examine the inhibitory effects of CO on haem-mediated PGRMC1 dimerization, SV-AUC analysis was carried out. RESULTS | |
| 30 34 haem chemical The peak corresponding to the haem/PGRMC1 dimer was detected under reducing conditions in the presence of dithionite (Supplementary Fig. 15, middle panel). RESULTS | |
| 35 41 PGRMC1 protein The peak corresponding to the haem/PGRMC1 dimer was detected under reducing conditions in the presence of dithionite (Supplementary Fig. 15, middle panel). RESULTS | |
| 42 47 dimer oligomeric_state The peak corresponding to the haem/PGRMC1 dimer was detected under reducing conditions in the presence of dithionite (Supplementary Fig. 15, middle panel). RESULTS | |
| 94 105 presence of protein_state The peak corresponding to the haem/PGRMC1 dimer was detected under reducing conditions in the presence of dithionite (Supplementary Fig. 15, middle panel). RESULTS | |
| 106 116 dithionite chemical The peak corresponding to the haem/PGRMC1 dimer was detected under reducing conditions in the presence of dithionite (Supplementary Fig. 15, middle panel). RESULTS | |
| 27 29 CO chemical Under these circumstances, CO application induced dissociation of the haem-mediated dimers of PGRMC1 to generate a peak of monomers (Supplementary Fig. 15, lower panel). RESULTS | |
| 70 74 haem chemical Under these circumstances, CO application induced dissociation of the haem-mediated dimers of PGRMC1 to generate a peak of monomers (Supplementary Fig. 15, lower panel). RESULTS | |
| 84 90 dimers oligomeric_state Under these circumstances, CO application induced dissociation of the haem-mediated dimers of PGRMC1 to generate a peak of monomers (Supplementary Fig. 15, lower panel). RESULTS | |
| 94 100 PGRMC1 protein Under these circumstances, CO application induced dissociation of the haem-mediated dimers of PGRMC1 to generate a peak of monomers (Supplementary Fig. 15, lower panel). RESULTS | |
| 123 131 monomers oligomeric_state Under these circumstances, CO application induced dissociation of the haem-mediated dimers of PGRMC1 to generate a peak of monomers (Supplementary Fig. 15, lower panel). RESULTS | |
| 76 82 PGRMC1 protein These observations raised the transition model for structural regulation of PGRMC1 in response to haem (Fig. 3d). RESULTS | |
| 98 102 haem chemical These observations raised the transition model for structural regulation of PGRMC1 in response to haem (Fig. 3d). RESULTS | |
| 20 23 apo protein_state As mentioned above, apo-PGRMC1 exists as monomer. RESULTS | |
| 24 30 PGRMC1 protein As mentioned above, apo-PGRMC1 exists as monomer. RESULTS | |
| 41 48 monomer oligomeric_state As mentioned above, apo-PGRMC1 exists as monomer. RESULTS | |
| 16 20 haem chemical By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 30 32 Kd evidence By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 47 53 PGRMC1 protein By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 62 68 stable protein_state By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 69 74 dimer oligomeric_state By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 76 88 dimerization oligomeric_state By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 89 91 Kd evidence By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 115 123 stacking bond_interaction By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 140 148 surfaces site By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 173 177 haem chemical By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 196 203 monomer oligomeric_state By binding with haem (binding Kd=50 nmol l−1), PGRMC1 forms a stable dimer (dimerization Kd<<3.5 μmol l−1) through stacking of the two open surfaces of the five-coordinated haem molecules in each monomer. RESULTS | |
| 0 2 CO chemical CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 35 39 haem chemical CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 49 54 dimer oligomeric_state CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 58 64 PGRMC1 protein CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 89 112 haem-stacking interface site CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 150 164 CO-haem-PGRMC1 complex_assembly CO induces the dissociation of the haem-mediated dimer of PGRMC1 by interfering with the haem-stacking interface via formation of the six-coordinated CO-haem-PGRMC1 complex. RESULTS | |
| 81 87 PGRMC1 protein Such a dynamic structural regulation led us to further examine the regulation of PGRMC1 functions in cancer cells. RESULTS | |
| 0 6 PGRMC1 protein PGRMC1 dimerization is required for binding to EGFR RESULTS | |
| 7 19 dimerization oligomeric_state PGRMC1 dimerization is required for binding to EGFR RESULTS | |
| 47 51 EGFR protein_type PGRMC1 dimerization is required for binding to EGFR RESULTS | |
| 8 14 PGRMC1 protein Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 41 45 EGFR protein_type Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 110 114 haem chemical Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 125 137 dimerization oligomeric_state Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 141 147 PGRMC1 protein Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 172 176 EGFR protein_type Because PGRMC1 is known to interact with EGFR and to accelerate tumour progression, we examined the effect of haem-dependent dimerization of PGRMC1 on its interaction with EGFR by using purified proteins. RESULTS | |
| 25 41 cytosolic domain structure_element As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 45 54 wild-type protein_state As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 55 61 PGRMC1 protein As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 75 80 Y113F mutant As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 81 87 mutant protein_state As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 114 118 EGFR protein_type As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 124 128 haem chemical As shown in Fig. 4a, the cytosolic domain of wild-type PGRMC1, but not the Y113F mutant, interacted with purified EGFR in a haem-dependent manner. RESULTS | |
| 38 47 ruthenium chemical This interaction was disrupted by the ruthenium-based CO-releasing molecule, CORM3, but not by RuCl3 as a control reagent (Fig. 4b). RESULTS | |
| 54 56 CO chemical This interaction was disrupted by the ruthenium-based CO-releasing molecule, CORM3, but not by RuCl3 as a control reagent (Fig. 4b). RESULTS | |
| 77 82 CORM3 chemical This interaction was disrupted by the ruthenium-based CO-releasing molecule, CORM3, but not by RuCl3 as a control reagent (Fig. 4b). RESULTS | |
| 95 100 RuCl3 chemical This interaction was disrupted by the ruthenium-based CO-releasing molecule, CORM3, but not by RuCl3 as a control reagent (Fig. 4b). RESULTS | |
| 58 64 PGRMC1 protein We further analysed the intracellular interaction between PGRMC1 and EGFR. RESULTS | |
| 69 73 EGFR protein_type We further analysed the intracellular interaction between PGRMC1 and EGFR. RESULTS | |
| 0 11 FLAG-tagged protein_state FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 12 18 PGRMC1 protein FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 19 40 ectopically expressed experimental_method FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 44 49 human species FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 80 98 immunoprecipitated experimental_method FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 128 149 co-immunoprecipitated experimental_method FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 150 154 EGFR protein_type FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 159 169 endogenous protein_state FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 170 176 PGRMC1 protein FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 193 199 PGRMC1 protein FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 217 233 Western blotting experimental_method FLAG-tagged PGRMC1 ectopically expressed in human colon cancer HCT116 cells was immunoprecipitated with anti-FLAG antibody, and co-immunoprecipitated EGFR and endogenous PGRMC1 binding to FLAG-PGRMC1 were detected by Western blotting (Fig. 4c). RESULTS | |
| 4 9 C129S mutant The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 10 16 mutant protein_state The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 20 26 PGRMC1 protein The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 48 58 endogenous protein_state The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 59 65 PGRMC1 protein The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 70 74 EGFR protein_type The C129S mutant of PGRMC1 also interacted with endogenous PGRMC1 and EGFR (Supplementary Fig. 16). RESULTS | |
| 8 19 FLAG-tagged protein_state Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 20 29 wild-type protein_state Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 30 36 PGRMC1 protein Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 53 63 endogenous protein_state Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 64 70 PGRMC1 protein Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 75 79 EGFR protein_type Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 85 90 Y113F mutant Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 91 97 mutant protein_state Whereas FLAG-tagged wild-type PGRMC1 interacted with endogenous PGRMC1 and EGFR, the Y113F mutant did not. RESULTS | |
| 31 46 succinylacetone chemical We also examined the effect of succinylacetone (SA), an inhibitor of haem biosynthesis (Fig. 4d). RESULTS | |
| 48 50 SA chemical We also examined the effect of succinylacetone (SA), an inhibitor of haem biosynthesis (Fig. 4d). RESULTS | |
| 69 73 haem chemical We also examined the effect of succinylacetone (SA), an inhibitor of haem biosynthesis (Fig. 4d). RESULTS | |
| 13 15 SA chemical As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 30 37 reduced protein_state As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 38 44 PGRMC1 protein As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 45 57 dimerization oligomeric_state As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 83 87 EGFR protein_type As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 115 119 haem chemical As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 129 141 dimerization oligomeric_state As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 145 151 PGMRC1 protein As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 183 187 EGFR protein_type As expected, SA significantly reduced PGRMC1 dimerization and its interaction with EGFR (Fig. 4e), indicating that haem-mediated dimerization of PGMRC1 is critical for its binding to EGFR. RESULTS | |
| 0 6 PGRMC1 protein PGRMC1 dimer facilitates EGFR-mediated cancer growth RESULTS | |
| 7 12 dimer oligomeric_state PGRMC1 dimer facilitates EGFR-mediated cancer growth RESULTS | |
| 25 29 EGFR protein_type PGRMC1 dimer facilitates EGFR-mediated cancer growth RESULTS | |
| 53 59 PGRMC1 protein Next, we investigated the functional significance of PGRMC1 dimerization in EGFR signaling. RESULTS | |
| 60 72 dimerization oligomeric_state Next, we investigated the functional significance of PGRMC1 dimerization in EGFR signaling. RESULTS | |
| 76 80 EGFR protein_type Next, we investigated the functional significance of PGRMC1 dimerization in EGFR signaling. RESULTS | |
| 0 3 EGF protein_type EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 12 28 phosphorylations ptm EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 32 36 EGFR protein_type EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 64 67 AKT protein_type EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 72 75 ERK protein_type EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 94 100 PGRMC1 protein EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 101 110 knockdown protein_state EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 112 121 PGRMC1-KD mutant EGF-induced phosphorylations of EGFR and its downstream targets AKT and ERK were decreased by PGRMC1 knockdown (PGRMC1-KD) (Fig. 4f). RESULTS | |
| 11 15 EGFR protein_type Similarly, EGFR signaling was suppressed by treatment of HCT116 cells with SA (Fig. 4g) or CORM3 (Fig. 4h). RESULTS | |
| 75 77 SA chemical Similarly, EGFR signaling was suppressed by treatment of HCT116 cells with SA (Fig. 4g) or CORM3 (Fig. 4h). RESULTS | |
| 91 96 CORM3 chemical Similarly, EGFR signaling was suppressed by treatment of HCT116 cells with SA (Fig. 4g) or CORM3 (Fig. 4h). RESULTS | |
| 29 33 haem chemical These results suggested that haem-mediated dimerization of PGRMC1 is critical for EGFR signaling. RESULTS | |
| 43 55 dimerization oligomeric_state These results suggested that haem-mediated dimerization of PGRMC1 is critical for EGFR signaling. RESULTS | |
| 59 65 PGRMC1 protein These results suggested that haem-mediated dimerization of PGRMC1 is critical for EGFR signaling. RESULTS | |
| 82 86 EGFR protein_type These results suggested that haem-mediated dimerization of PGRMC1 is critical for EGFR signaling. RESULTS | |
| 39 48 dimerized protein_state To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 57 63 PGRMC1 protein To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 102 108 PGRMC1 protein To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 109 137 knockdown-rescue experiments experimental_method To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 144 155 FLAG-tagged protein_state To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 156 165 wild-type protein_state To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 170 175 Y113F mutant To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 176 182 PGRMC1 protein To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 183 201 expression vectors experimental_method To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 212 228 silent mutations experimental_method To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 234 244 introduced experimental_method To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 286 291 shRNA chemical To further investigate the role of the dimerized form of PGRMC1 in cancer proliferation, we performed PGRMC1 knockdown-rescue experiments using FLAG-tagged wild-type and Y113F PGRMC1 expression vectors, in which silent mutations were introduced into the nucleotide sequence targeted by shRNA (Fig. 5a). RESULTS | |
| 56 69 knocking down experimental_method While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 70 76 PGRMC1 protein While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 78 87 PGRMC1-KD mutant While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 121 125 EGFR protein_type While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 136 145 erlotinib chemical While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 214 227 co-expression experimental_method While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 231 246 shRNA-resistant protein_state While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 247 256 wild-type protein_state While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 257 263 PGRMC1 protein While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 279 284 Y113F mutant While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 285 291 mutant protein_state While proliferation of HCT116 cells was not affected by knocking down PGRMC1, PGRMC1-KD cells were more sensitive to the EGFR inhibitor erlotinib than control HCT116 cells, and the knockdown effect was reversed by co-expression of shRNA-resistant wild-type PGRMC1 but not of the Y113F mutant (Fig. 5b). RESULTS | |
| 46 52 shRNAs chemical Chemosensitivity enhancement by two different shRNAs to PGRMC1 was seen also in HCT116 cells and human hepatoma HuH7 cells (Supplementary Fig. 17). RESULTS | |
| 56 62 PGRMC1 protein Chemosensitivity enhancement by two different shRNAs to PGRMC1 was seen also in HCT116 cells and human hepatoma HuH7 cells (Supplementary Fig. 17). RESULTS | |
| 97 102 human species Chemosensitivity enhancement by two different shRNAs to PGRMC1 was seen also in HCT116 cells and human hepatoma HuH7 cells (Supplementary Fig. 17). RESULTS | |
| 13 22 PGRMC1-KD mutant Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 116 129 co-expression experimental_method Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 133 142 wild-type protein_state Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 143 149 PGRMC1 protein Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 165 170 Y113F mutant Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 171 177 mutant protein_state Furthermore, PGRMC1-KD inhibited spheroid formation of HCT116 cells in culture, and this inhibition was reversed by co-expression of wild-type PGRMC1 but not of the Y113F mutant (Fig. 5c and Supplementary Fig. 18). RESULTS | |
| 6 12 PGRMC1 protein Thus, PGRMC1 dimerization is important for cancer cell proliferation and chemoresistance. RESULTS | |
| 13 25 dimerization oligomeric_state Thus, PGRMC1 dimerization is important for cancer cell proliferation and chemoresistance. RESULTS | |
| 24 30 PGRMC1 protein We examined the role of PGRMC1 in metastatic progression by xenograft transplantation assays using super-immunodeficient NOD/scid/γnull (NOG) mice. RESULTS | |
| 60 92 xenograft transplantation assays experimental_method We examined the role of PGRMC1 in metastatic progression by xenograft transplantation assays using super-immunodeficient NOD/scid/γnull (NOG) mice. RESULTS | |
| 15 41 intra-splenic implantation experimental_method Ten days after intra-splenic implantation of HCT116 cells that were genetically tagged with a fluorescent protein Venus, the group implanted with PGRMC1-KD cells showed a significant decrease of liver metastasis in comparison with the control group (Fig. 5d). RESULTS | |
| 146 155 PGRMC1-KD mutant Ten days after intra-splenic implantation of HCT116 cells that were genetically tagged with a fluorescent protein Venus, the group implanted with PGRMC1-KD cells showed a significant decrease of liver metastasis in comparison with the control group (Fig. 5d). RESULTS | |
| 15 21 PGRMC1 protein Interaction of PGRMC1 dimer with cytochromes P450 RESULTS | |
| 22 27 dimer oligomeric_state Interaction of PGRMC1 dimer with cytochromes P450 RESULTS | |
| 33 49 cytochromes P450 protein_type Interaction of PGRMC1 dimer with cytochromes P450 RESULTS | |
| 6 12 PGRMC1 protein Since PGRMC1 has been shown to interact with cytochromes P450 (ref), we investigated whether the haem-mediated dimerization of PGRMC1 is necessary for their interactions. RESULTS | |
| 45 61 cytochromes P450 protein_type Since PGRMC1 has been shown to interact with cytochromes P450 (ref), we investigated whether the haem-mediated dimerization of PGRMC1 is necessary for their interactions. RESULTS | |
| 97 101 haem chemical Since PGRMC1 has been shown to interact with cytochromes P450 (ref), we investigated whether the haem-mediated dimerization of PGRMC1 is necessary for their interactions. RESULTS | |
| 111 123 dimerization oligomeric_state Since PGRMC1 has been shown to interact with cytochromes P450 (ref), we investigated whether the haem-mediated dimerization of PGRMC1 is necessary for their interactions. RESULTS | |
| 127 133 PGRMC1 protein Since PGRMC1 has been shown to interact with cytochromes P450 (ref), we investigated whether the haem-mediated dimerization of PGRMC1 is necessary for their interactions. RESULTS | |
| 12 18 CYP1A2 protein Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 23 29 CYP3A4 protein Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 76 89 cytochrome b5 protein_type Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 94 119 cytochrome P450 reductase protein Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 139 155 cytochromes P450 protein_type Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 173 182 wild-type protein_state Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 183 189 PGRMC1 protein Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 208 213 Y113F mutant Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 214 220 mutant protein_state Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 227 231 haem chemical Recombinant CYP1A2 and CYP3A4 including a microsomal formulation containing cytochrome b5 and cytochrome P450 reductase, drug-metabolizing cytochromes P450, interacted with wild-type PGRMC1, but not with the Y113F mutant, in a haem-dependent manner (Fig. 6a,b). RESULTS | |
| 29 35 PGRMC1 protein Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 41 47 CYP1A2 protein Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 63 68 CORM3 chemical Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 122 128 PGRMC1 protein Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 129 141 dimerization oligomeric_state Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 180 196 cytochromes P450 protein_type Moreover, the interaction of PGRMC1 with CYP1A2 was blocked by CORM3 under reducing conditions (Fig. 6c), indicating that PGRMC1 dimerization is necessary for its interaction with cytochromes P450. RESULTS | |
| 0 11 Doxorubicin chemical Doxorubicin is an anti-cancer reagent that is metabolized into inactive doxorubicinol by CYP2D6 and CYP3A4 (Fig. 6d). RESULTS | |
| 72 85 doxorubicinol chemical Doxorubicin is an anti-cancer reagent that is metabolized into inactive doxorubicinol by CYP2D6 and CYP3A4 (Fig. 6d). RESULTS | |
| 89 95 CYP2D6 protein Doxorubicin is an anti-cancer reagent that is metabolized into inactive doxorubicinol by CYP2D6 and CYP3A4 (Fig. 6d). RESULTS | |
| 100 106 CYP3A4 protein Doxorubicin is an anti-cancer reagent that is metabolized into inactive doxorubicinol by CYP2D6 and CYP3A4 (Fig. 6d). RESULTS | |
| 0 9 PGRMC1-KD mutant PGRMC1-KD significantly suppressed the conversion of doxorubicin to doxorubicinol (Fig. 6d) and increased sensitivity to doxorubicin (Fig. 6e). RESULTS | |
| 53 64 doxorubicin chemical PGRMC1-KD significantly suppressed the conversion of doxorubicin to doxorubicinol (Fig. 6d) and increased sensitivity to doxorubicin (Fig. 6e). RESULTS | |
| 68 81 doxorubicinol chemical PGRMC1-KD significantly suppressed the conversion of doxorubicin to doxorubicinol (Fig. 6d) and increased sensitivity to doxorubicin (Fig. 6e). RESULTS | |
| 121 132 doxorubicin chemical PGRMC1-KD significantly suppressed the conversion of doxorubicin to doxorubicinol (Fig. 6d) and increased sensitivity to doxorubicin (Fig. 6e). RESULTS | |
| 9 20 doxorubicin chemical Enhanced doxorubicin sensitivity was modestly but significantly induced by PGRMC1-KD. RESULTS | |
| 75 84 PGRMC1-KD mutant Enhanced doxorubicin sensitivity was modestly but significantly induced by PGRMC1-KD. RESULTS | |
| 28 41 co-expression experimental_method This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 49 58 wild-type protein_state This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 59 65 PGRMC1 protein This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 81 86 Y113F mutant This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 87 93 mutant protein_state This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 111 117 PGRMC1 protein This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 127 138 doxorubicin chemical This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 202 218 cytochromes P450 protein_type This effect was reversed by co-expression of the wild-type PGRMC1 but not of the Y113F mutant, suggesting that PGRMC1 enhances doxorubicin resistance of cancer cells by facilitating its degradation via cytochromes P450. RESULTS | |
| 53 59 PGRMC1 protein To gain further insight into the interaction between PGRMC1 and cytochromes P450, surface plasmon resonance analyses were conducted using recombinant CYP51 and PGRMC1. RESULTS | |
| 64 80 cytochromes P450 protein_type To gain further insight into the interaction between PGRMC1 and cytochromes P450, surface plasmon resonance analyses were conducted using recombinant CYP51 and PGRMC1. RESULTS | |
| 82 116 surface plasmon resonance analyses experimental_method To gain further insight into the interaction between PGRMC1 and cytochromes P450, surface plasmon resonance analyses were conducted using recombinant CYP51 and PGRMC1. RESULTS | |
| 150 155 CYP51 protein To gain further insight into the interaction between PGRMC1 and cytochromes P450, surface plasmon resonance analyses were conducted using recombinant CYP51 and PGRMC1. RESULTS | |
| 160 166 PGRMC1 protein To gain further insight into the interaction between PGRMC1 and cytochromes P450, surface plasmon resonance analyses were conducted using recombinant CYP51 and PGRMC1. RESULTS | |
| 48 54 PGRMC1 protein This was based on a previous study showing that PGRMC1 binds to CYP51 and enhances cholesterol biosynthesis by CYP51 (refs). RESULTS | |
| 64 69 CYP51 protein This was based on a previous study showing that PGRMC1 binds to CYP51 and enhances cholesterol biosynthesis by CYP51 (refs). RESULTS | |
| 111 116 CYP51 protein This was based on a previous study showing that PGRMC1 binds to CYP51 and enhances cholesterol biosynthesis by CYP51 (refs). RESULTS | |
| 0 5 CYP51 protein CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 22 28 PGRMC1 protein CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 72 83 presence of protein_state CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 84 88 haem chemical CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 105 112 absence protein_state CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 173 177 haem chemical CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 188 200 dimerization oligomeric_state CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 204 210 PGRMC1 protein CYP51 interacted with PGRMC1 in a concentration-dependent manner in the presence of haem, but not in its absence (Supplementary Fig. 19), suggesting the requirement for the haem-dependent dimerization of PGRMC1. RESULTS | |
| 4 6 Kd evidence The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 16 22 PGRMC1 protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 34 39 CYP51 protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 101 105 haem chemical The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 124 149 cytochrome P450 reductase protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 154 165 neuroglobin protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 166 170 Gαi1 protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 195 199 haem chemical The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 210 216 PGRMC1 protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 234 239 CYP51 protein The Kd value of PGRMC1 binding to CYP51 was in a micromolar range and comparable with those of other haem proteins, such as cytochrome P450 reductase and neuroglobin/Gαi1 (ref.), suggesting that haem-dependent PGRMC1 interaction with CYP51 is biologically relevant. RESULTS | |
| 30 36 PGRMC1 protein In this study, we showed that PGRMC1 dimerizes by stacking interactions of haem molecules from each monomer. DISCUSS | |
| 37 46 dimerizes oligomeric_state In this study, we showed that PGRMC1 dimerizes by stacking interactions of haem molecules from each monomer. DISCUSS | |
| 50 71 stacking interactions bond_interaction In this study, we showed that PGRMC1 dimerizes by stacking interactions of haem molecules from each monomer. DISCUSS | |
| 75 79 haem chemical In this study, we showed that PGRMC1 dimerizes by stacking interactions of haem molecules from each monomer. DISCUSS | |
| 100 107 monomer oligomeric_state In this study, we showed that PGRMC1 dimerizes by stacking interactions of haem molecules from each monomer. DISCUSS | |
| 37 78 translationally-controlled tumour protein protein_type Recently, Lucas et al. reported that translationally-controlled tumour protein was dimerized by binding with haem, but its structural basis remains unclear. DISCUSS | |
| 83 92 dimerized protein_state Recently, Lucas et al. reported that translationally-controlled tumour protein was dimerized by binding with haem, but its structural basis remains unclear. DISCUSS | |
| 109 113 haem chemical Recently, Lucas et al. reported that translationally-controlled tumour protein was dimerized by binding with haem, but its structural basis remains unclear. DISCUSS | |
| 88 106 haem–haem stacking bond_interaction This is the report showing crystallographic evidence that indicates roles of the direct haem–haem stacking in haem-mediated dimerization in eukaryotes, although a few examples are known in bacteria. DISCUSS | |
| 110 114 haem chemical This is the report showing crystallographic evidence that indicates roles of the direct haem–haem stacking in haem-mediated dimerization in eukaryotes, although a few examples are known in bacteria. DISCUSS | |
| 124 136 dimerization oligomeric_state This is the report showing crystallographic evidence that indicates roles of the direct haem–haem stacking in haem-mediated dimerization in eukaryotes, although a few examples are known in bacteria. DISCUSS | |
| 140 150 eukaryotes taxonomy_domain This is the report showing crystallographic evidence that indicates roles of the direct haem–haem stacking in haem-mediated dimerization in eukaryotes, although a few examples are known in bacteria. DISCUSS | |
| 189 197 bacteria taxonomy_domain This is the report showing crystallographic evidence that indicates roles of the direct haem–haem stacking in haem-mediated dimerization in eukaryotes, although a few examples are known in bacteria. DISCUSS | |
| 0 19 Sequence alignments experimental_method Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 30 51 haem-binding residues site Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 53 59 Tyr113 residue_name_number Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 61 67 Tyr107 residue_name_number Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 69 75 Lys163 residue_name_number Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 80 86 Tyr164 residue_name_number Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 91 97 PGRMC1 protein Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 102 111 conserved protein_state Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 118 122 MAPR protein_type Sequence alignments show that haem-binding residues (Tyr113, Tyr107, Lys163 and Tyr164) in PGRMC1 are conserved among MAPR proteins (Supplementary Fig. 5). DISCUSS | |
| 26 30 Y113 residue_name_number In the current study, the Y113 residue plays a crucial role for the haem-dependent dimerization of PGRMC1 and resultant regulation of cancer proliferation and chemoresistance (Figs 5c and 6e). DISCUSS | |
| 68 72 haem chemical In the current study, the Y113 residue plays a crucial role for the haem-dependent dimerization of PGRMC1 and resultant regulation of cancer proliferation and chemoresistance (Figs 5c and 6e). DISCUSS | |
| 83 95 dimerization oligomeric_state In the current study, the Y113 residue plays a crucial role for the haem-dependent dimerization of PGRMC1 and resultant regulation of cancer proliferation and chemoresistance (Figs 5c and 6e). DISCUSS | |
| 99 105 PGRMC1 protein In the current study, the Y113 residue plays a crucial role for the haem-dependent dimerization of PGRMC1 and resultant regulation of cancer proliferation and chemoresistance (Figs 5c and 6e). DISCUSS | |
| 10 14 Y113 residue_name_number Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 51 66 consensus motif structure_element Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 70 85 phosphorylation ptm Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 89 105 tyrosine kinases protein_type Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 114 117 Abl protein_type Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 122 125 Lck protein_type Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 151 165 phosphorylated protein_state Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 166 170 Y113 residue_name_number Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 201 207 ESI-MS experimental_method Since the Y113 residue is involved in the putative consensus motif of phosphorylation by tyrosine kinases such as Abl and Lck, we investigated whether phosphorylated Y113 is present in HCT116 cells by ESI-MS analysis. DISCUSS | |
| 38 44 PGRMC1 protein Recently, Peluso et al. reported that PGRMC1 binds to PGRMC2, suggesting that MAPR family members may also undergo haem-mediated heterodimerization. DISCUSS | |
| 54 60 PGRMC2 protein Recently, Peluso et al. reported that PGRMC1 binds to PGRMC2, suggesting that MAPR family members may also undergo haem-mediated heterodimerization. DISCUSS | |
| 78 82 MAPR protein_type Recently, Peluso et al. reported that PGRMC1 binds to PGRMC2, suggesting that MAPR family members may also undergo haem-mediated heterodimerization. DISCUSS | |
| 115 119 haem chemical Recently, Peluso et al. reported that PGRMC1 binds to PGRMC2, suggesting that MAPR family members may also undergo haem-mediated heterodimerization. DISCUSS | |
| 19 23 haem chemical We showed that the haem-mediated dimer of PGRMC1 enables interaction with different subclasses of cytochromes P450 (CYP) (Fig. 6). DISCUSS | |
| 33 38 dimer oligomeric_state We showed that the haem-mediated dimer of PGRMC1 enables interaction with different subclasses of cytochromes P450 (CYP) (Fig. 6). DISCUSS | |
| 42 48 PGRMC1 protein We showed that the haem-mediated dimer of PGRMC1 enables interaction with different subclasses of cytochromes P450 (CYP) (Fig. 6). DISCUSS | |
| 98 114 cytochromes P450 protein_type We showed that the haem-mediated dimer of PGRMC1 enables interaction with different subclasses of cytochromes P450 (CYP) (Fig. 6). DISCUSS | |
| 116 119 CYP protein_type We showed that the haem-mediated dimer of PGRMC1 enables interaction with different subclasses of cytochromes P450 (CYP) (Fig. 6). DISCUSS | |
| 21 27 PGRMC1 protein While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 65 70 CYP51 protein While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 100 105 yeast taxonomy_domain While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 110 115 human species While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 171 174 CYP protein_type While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 203 209 PGRMC1 protein While the effects of PGRMC1 on cholesterol synthesis mediated by CYP51 have been well documented in yeast and human cells, it has not been clear whether drug-metabolizing CYP activities are regulated by PGRMC1. DISCUSS | |
| 42 48 PGRMC1 protein Szczesna-Skorupa and Kemper reported that PGRMC1 exhibited an inhibitory effect on CYP3A4 drug metabolizing activity by competitively binding with cytochrome P450 reductase (CPR) in HEK293 or HepG2 cells. DISCUSS | |
| 83 89 CYP3A4 protein Szczesna-Skorupa and Kemper reported that PGRMC1 exhibited an inhibitory effect on CYP3A4 drug metabolizing activity by competitively binding with cytochrome P450 reductase (CPR) in HEK293 or HepG2 cells. DISCUSS | |
| 147 172 cytochrome P450 reductase protein Szczesna-Skorupa and Kemper reported that PGRMC1 exhibited an inhibitory effect on CYP3A4 drug metabolizing activity by competitively binding with cytochrome P450 reductase (CPR) in HEK293 or HepG2 cells. DISCUSS | |
| 174 177 CPR protein Szczesna-Skorupa and Kemper reported that PGRMC1 exhibited an inhibitory effect on CYP3A4 drug metabolizing activity by competitively binding with cytochrome P450 reductase (CPR) in HEK293 or HepG2 cells. DISCUSS | |
| 44 50 PGRMC1 protein On the other hand, Oda et al. reported that PGRMC1 had no effect to CYP2E1 and CYP3A4 activities in HepG2 cell. DISCUSS | |
| 68 74 CYP2E1 protein On the other hand, Oda et al. reported that PGRMC1 had no effect to CYP2E1 and CYP3A4 activities in HepG2 cell. DISCUSS | |
| 79 85 CYP3A4 protein On the other hand, Oda et al. reported that PGRMC1 had no effect to CYP2E1 and CYP3A4 activities in HepG2 cell. DISCUSS | |
| 33 39 PGRMC1 protein Several other groups showed that PGRMC1 enhanced chemoresistance in several cancer cells such as uterine sarcoma, breast cancer, endometrial tumour and ovarian cancer; however, no evidence of PGRMC1-dependent regulation of CYP activity was provided. DISCUSS | |
| 192 198 PGRMC1 protein Several other groups showed that PGRMC1 enhanced chemoresistance in several cancer cells such as uterine sarcoma, breast cancer, endometrial tumour and ovarian cancer; however, no evidence of PGRMC1-dependent regulation of CYP activity was provided. DISCUSS | |
| 223 226 CYP protein_type Several other groups showed that PGRMC1 enhanced chemoresistance in several cancer cells such as uterine sarcoma, breast cancer, endometrial tumour and ovarian cancer; however, no evidence of PGRMC1-dependent regulation of CYP activity was provided. DISCUSS | |
| 24 30 PGRMC1 protein Our results showed that PGRMC1 contributes to enhancement of the doxorubicin metabolism, which is mediated by CYP2D6 or CYP3A4 in human colon cancer HCT116 cells (Fig. 6d). DISCUSS | |
| 65 76 doxorubicin chemical Our results showed that PGRMC1 contributes to enhancement of the doxorubicin metabolism, which is mediated by CYP2D6 or CYP3A4 in human colon cancer HCT116 cells (Fig. 6d). DISCUSS | |
| 110 116 CYP2D6 protein Our results showed that PGRMC1 contributes to enhancement of the doxorubicin metabolism, which is mediated by CYP2D6 or CYP3A4 in human colon cancer HCT116 cells (Fig. 6d). DISCUSS | |
| 120 126 CYP3A4 protein Our results showed that PGRMC1 contributes to enhancement of the doxorubicin metabolism, which is mediated by CYP2D6 or CYP3A4 in human colon cancer HCT116 cells (Fig. 6d). DISCUSS | |
| 130 135 human species Our results showed that PGRMC1 contributes to enhancement of the doxorubicin metabolism, which is mediated by CYP2D6 or CYP3A4 in human colon cancer HCT116 cells (Fig. 6d). DISCUSS | |
| 45 48 CYP protein_type While the effects of structural diversity of CYP family proteins and interactions with different xenobiotic substrates should further be examined, the current results suggest that the interaction of drug-metabolizing CYPs with the haem-mediated dimer of PGRMC1 plays a crucial role in regulating their activities. DISCUSS | |
| 217 221 CYPs protein_type While the effects of structural diversity of CYP family proteins and interactions with different xenobiotic substrates should further be examined, the current results suggest that the interaction of drug-metabolizing CYPs with the haem-mediated dimer of PGRMC1 plays a crucial role in regulating their activities. DISCUSS | |
| 231 235 haem chemical While the effects of structural diversity of CYP family proteins and interactions with different xenobiotic substrates should further be examined, the current results suggest that the interaction of drug-metabolizing CYPs with the haem-mediated dimer of PGRMC1 plays a crucial role in regulating their activities. DISCUSS | |
| 245 250 dimer oligomeric_state While the effects of structural diversity of CYP family proteins and interactions with different xenobiotic substrates should further be examined, the current results suggest that the interaction of drug-metabolizing CYPs with the haem-mediated dimer of PGRMC1 plays a crucial role in regulating their activities. DISCUSS | |
| 254 260 PGRMC1 protein While the effects of structural diversity of CYP family proteins and interactions with different xenobiotic substrates should further be examined, the current results suggest that the interaction of drug-metabolizing CYPs with the haem-mediated dimer of PGRMC1 plays a crucial role in regulating their activities. DISCUSS | |
| 15 19 haem chemical We showed that haem-mediated dimerization of PGRMC1 enhances proliferation and chemoresistance of cancer cells through binding to and regulating EGFR and cytochromes P450 (illustrated in Fig. 7). DISCUSS | |
| 29 41 dimerization oligomeric_state We showed that haem-mediated dimerization of PGRMC1 enhances proliferation and chemoresistance of cancer cells through binding to and regulating EGFR and cytochromes P450 (illustrated in Fig. 7). DISCUSS | |
| 45 51 PGRMC1 protein We showed that haem-mediated dimerization of PGRMC1 enhances proliferation and chemoresistance of cancer cells through binding to and regulating EGFR and cytochromes P450 (illustrated in Fig. 7). DISCUSS | |
| 145 149 EGFR protein_type We showed that haem-mediated dimerization of PGRMC1 enhances proliferation and chemoresistance of cancer cells through binding to and regulating EGFR and cytochromes P450 (illustrated in Fig. 7). DISCUSS | |
| 154 170 cytochromes P450 protein_type We showed that haem-mediated dimerization of PGRMC1 enhances proliferation and chemoresistance of cancer cells through binding to and regulating EGFR and cytochromes P450 (illustrated in Fig. 7). DISCUSS | |
| 10 31 haem-binding affinity evidence Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 35 41 PGRMC1 protein Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 65 77 constitutive protein_state Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 78 99 haem-binding proteins protein_type Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 108 117 myoglobin protein Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 119 125 PGMRC1 protein Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 161 164 apo protein_state Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 165 172 monomer oligomeric_state Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 177 187 haem-bound protein_state Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 188 193 dimer oligomeric_state Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 244 248 haem chemical Since the haem-binding affinity of PGRMC1 is lower than those of constitutive haem-binding proteins such as myoglobin, PGMRC1 is probably interconverted between apo-monomer and haem-bound dimer forms in response to changes in the intracellular haem concentration. DISCUSS | |
| 67 71 haem chemical Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 191 195 haem chemical Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 249 267 fumarate hydratase protein_type Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 297 309 succinyl CoA chemical Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 327 331 haem chemical Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 369 373 haem chemical Considering microenvironments in and around malignant tumours, the haem concentration in cancer cells is likely to be elevated through multiple mechanisms, such as (i) an increased intake of haem, (ii) mutation of enzymes in TCA cycle (for example, fumarate hydratase) that increases the level of succinyl CoA, a substrate for haem biosynthesis and (iii) metastasis to haem-rich organs such as liver, brain and bone marrow. DISCUSS | |
| 220 224 haem chemical Moreover, exposure of cancer cells to stimuli such as hypoxia, radiation and chemotherapy causes cell damages and leads to protein degradation, resulting in increased levels of TCA cycle intermediates and in an enhanced haem biosynthesis. DISCUSS | |
| 29 33 haem chemical On the other hand, excessive haem induces HO-1, the enzyme that oxidatively degrades haem and generates CO. DISCUSS | |
| 42 46 HO-1 protein On the other hand, excessive haem induces HO-1, the enzyme that oxidatively degrades haem and generates CO. DISCUSS | |
| 85 89 haem chemical On the other hand, excessive haem induces HO-1, the enzyme that oxidatively degrades haem and generates CO. DISCUSS | |
| 104 106 CO chemical On the other hand, excessive haem induces HO-1, the enzyme that oxidatively degrades haem and generates CO. DISCUSS | |
| 6 10 HO-1 protein Thus, HO-1 induction in cancer cells may inhibit the haem-mediated dimerization of PGRMC1 through the production of CO and thereby suppress tumour progression. DISCUSS | |
| 53 57 haem chemical Thus, HO-1 induction in cancer cells may inhibit the haem-mediated dimerization of PGRMC1 through the production of CO and thereby suppress tumour progression. DISCUSS | |
| 67 79 dimerization oligomeric_state Thus, HO-1 induction in cancer cells may inhibit the haem-mediated dimerization of PGRMC1 through the production of CO and thereby suppress tumour progression. DISCUSS | |
| 83 89 PGRMC1 protein Thus, HO-1 induction in cancer cells may inhibit the haem-mediated dimerization of PGRMC1 through the production of CO and thereby suppress tumour progression. DISCUSS | |
| 116 118 CO chemical Thus, HO-1 induction in cancer cells may inhibit the haem-mediated dimerization of PGRMC1 through the production of CO and thereby suppress tumour progression. DISCUSS | |
| 50 54 HO-1 protein This idea is consistent with the observation that HO-1 induction or CO inhibits tumour growth. DISCUSS | |
| 68 70 CO chemical This idea is consistent with the observation that HO-1 induction or CO inhibits tumour growth. DISCUSS | |
| 32 38 PGRMC1 protein Besides the regulatory roles of PGRMC1/Sigma-2 receptor in proliferation and chemoresistance in cancer cells (ref.), recent reports show that PGRMC1 is able to bind to amyloid beta oligomer to enhance its neurotoxicity. DISCUSS | |
| 39 46 Sigma-2 protein Besides the regulatory roles of PGRMC1/Sigma-2 receptor in proliferation and chemoresistance in cancer cells (ref.), recent reports show that PGRMC1 is able to bind to amyloid beta oligomer to enhance its neurotoxicity. DISCUSS | |
| 142 148 PGRMC1 protein Besides the regulatory roles of PGRMC1/Sigma-2 receptor in proliferation and chemoresistance in cancer cells (ref.), recent reports show that PGRMC1 is able to bind to amyloid beta oligomer to enhance its neurotoxicity. DISCUSS | |
| 168 180 amyloid beta protein Besides the regulatory roles of PGRMC1/Sigma-2 receptor in proliferation and chemoresistance in cancer cells (ref.), recent reports show that PGRMC1 is able to bind to amyloid beta oligomer to enhance its neurotoxicity. DISCUSS | |
| 181 189 oligomer oligomeric_state Besides the regulatory roles of PGRMC1/Sigma-2 receptor in proliferation and chemoresistance in cancer cells (ref.), recent reports show that PGRMC1 is able to bind to amyloid beta oligomer to enhance its neurotoxicity. DISCUSS | |
| 13 20 Sigma-2 protein Furthermore, Sigma-2 ligand-binding is decreased in transgenic amyloid beta deposition model APP/PS1 female mice. DISCUSS | |
| 48 54 PGRMC1 protein These results suggest a possible involvement of PGRMC1 in Alzheimer's disease. DISCUSS | |
| 13 17 haem chemical The roles of haem-dependent dimerization of PGRMC1 in the functional regulation of its target proteins deserve further studies to find evidence that therapeutic interventions to interfere with the function of the dimer may control varied disease conditions. DISCUSS | |
| 28 40 dimerization oligomeric_state The roles of haem-dependent dimerization of PGRMC1 in the functional regulation of its target proteins deserve further studies to find evidence that therapeutic interventions to interfere with the function of the dimer may control varied disease conditions. DISCUSS | |
| 44 50 PGRMC1 protein The roles of haem-dependent dimerization of PGRMC1 in the functional regulation of its target proteins deserve further studies to find evidence that therapeutic interventions to interfere with the function of the dimer may control varied disease conditions. DISCUSS | |
| 213 218 dimer oligomeric_state The roles of haem-dependent dimerization of PGRMC1 in the functional regulation of its target proteins deserve further studies to find evidence that therapeutic interventions to interfere with the function of the dimer may control varied disease conditions. DISCUSS | |
| 109 117 oligomer oligomeric_state Alzheimer's therapeutics targeting amyloid beta 1-42 oligomers II: Sigma-2/PGRMC1 receptors mediate Abeta 42 oligomer binding and synaptotoxicity REF | |
| 0 23 X-ray crystal structure evidence X-ray crystal structure of PGRMC1. FIG | |
| 27 33 PGRMC1 protein X-ray crystal structure of PGRMC1. FIG | |
| 21 27 PGRMC1 protein (a) Structure of the PGRMC1 dimer formed through stacked haems. FIG | |
| 28 33 dimer oligomeric_state (a) Structure of the PGRMC1 dimer formed through stacked haems. FIG | |
| 57 62 haems chemical (a) Structure of the PGRMC1 dimer formed through stacked haems. FIG | |
| 4 10 PGRMC1 protein Two PGRMC1 subunits (blue and green ribbons) dimerize via stacking of the haem molecules. FIG | |
| 11 19 subunits structure_element Two PGRMC1 subunits (blue and green ribbons) dimerize via stacking of the haem molecules. FIG | |
| 45 53 dimerize oligomeric_state Two PGRMC1 subunits (blue and green ribbons) dimerize via stacking of the haem molecules. FIG | |
| 58 66 stacking bond_interaction Two PGRMC1 subunits (blue and green ribbons) dimerize via stacking of the haem molecules. FIG | |
| 74 78 haem chemical Two PGRMC1 subunits (blue and green ribbons) dimerize via stacking of the haem molecules. FIG | |
| 4 8 Haem chemical (b) Haem coordination of PGRMC1 with Tyr113. FIG | |
| 9 21 coordination bond_interaction (b) Haem coordination of PGRMC1 with Tyr113. FIG | |
| 25 31 PGRMC1 protein (b) Haem coordination of PGRMC1 with Tyr113. FIG | |
| 37 43 Tyr113 residue_name_number (b) Haem coordination of PGRMC1 with Tyr113. FIG | |
| 14 20 PGRMC1 protein Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 32 45 cytochrome b5 protein_type Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 70 76 PGRMC1 protein Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 90 95 helix structure_element Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 101 108 147–163 residue_range Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 142 146 haem chemical Comparison of PGRMC1 (blue) and cytochrome b5 (yellow, ID: 3NER). (c) PGRMC1 has a longer helix (a.a.147–163), which is shifted away from the haem (arrow). FIG | |
| 0 6 PGRCM1 protein PGRCM1 is dimerized by binding with haem. FIG | |
| 10 19 dimerized protein_state PGRCM1 is dimerized by binding with haem. FIG | |
| 36 40 haem chemical PGRCM1 is dimerized by binding with haem. FIG | |
| 4 22 Mass spectrometric experimental_method (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 39 48 wild-type protein_state (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 50 52 wt protein_state (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 54 60 PGRMC1 protein (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 68 73 C129S mutant (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 74 80 mutant protein_state (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 88 96 presence protein_state (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 100 110 absence of protein_state (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 111 115 haem chemical (a) Mass spectrometric analyses of the wild-type (wt) PGRMC1 or the C129S mutant in the presence or absence of haem under non-denaturing condition. FIG | |
| 41 47 44–195 residue_range Both proteins had identical lengths (a.a.44–195). FIG | |
| 4 10 SV-AUC experimental_method (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 27 29 wt protein_state (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 30 36 PGRMC1 protein (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 45 50 C129S mutant (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 51 57 mutant protein_state (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 63 69 44–195 residue_range (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 78 86 presence protein_state (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 90 100 absence of protein_state (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 101 105 haem chemical (b) SV-AUC analyses of the wt-PGRMC1 and the C129S mutant (a.a.44–195) in the presence or absence of haem. FIG | |
| 0 6 SV-AUC experimental_method SV-AUC experiments were performed with 1.5 mg ml−1 of PGRMC1 proteins. FIG | |
| 54 60 PGRMC1 protein SV-AUC experiments were performed with 1.5 mg ml−1 of PGRMC1 proteins. FIG | |
| 20 45 sedimentation coefficient evidence The major peak with sedimentation coefficient S20,w of 1.9∼2.0 S (monomer) or 3.1 S (dimer) was detected. FIG | |
| 46 51 S20,w evidence The major peak with sedimentation coefficient S20,w of 1.9∼2.0 S (monomer) or 3.1 S (dimer) was detected. FIG | |
| 66 73 monomer oligomeric_state The major peak with sedimentation coefficient S20,w of 1.9∼2.0 S (monomer) or 3.1 S (dimer) was detected. FIG | |
| 85 90 dimer oligomeric_state The major peak with sedimentation coefficient S20,w of 1.9∼2.0 S (monomer) or 3.1 S (dimer) was detected. FIG | |
| 4 33 Difference absorption spectra evidence (c) Difference absorption spectra of PGRMC1 (a.a.44–195) titrated with haem (left panel). FIG | |
| 37 43 PGRMC1 protein (c) Difference absorption spectra of PGRMC1 (a.a.44–195) titrated with haem (left panel). FIG | |
| 49 55 44–195 residue_range (c) Difference absorption spectra of PGRMC1 (a.a.44–195) titrated with haem (left panel). FIG | |
| 57 70 titrated with experimental_method (c) Difference absorption spectra of PGRMC1 (a.a.44–195) titrated with haem (left panel). FIG | |
| 71 75 haem chemical (c) Difference absorption spectra of PGRMC1 (a.a.44–195) titrated with haem (left panel). FIG | |
| 4 19 titration curve evidence The titration curve of haem to PGRMC1 (right panel). FIG | |
| 23 27 haem chemical The titration curve of haem to PGRMC1 (right panel). FIG | |
| 31 37 PGRMC1 protein The titration curve of haem to PGRMC1 (right panel). FIG | |
| 4 25 absorbance difference evidence The absorbance difference at 400 nm is plotted against the haem concentration. FIG | |
| 59 63 haem chemical The absorbance difference at 400 nm is plotted against the haem concentration. FIG | |
| 0 15 Carbon monoxide chemical Carbon monoxide inhibits haem-dependent PGRMC1 dimerization. FIG | |
| 25 29 haem chemical Carbon monoxide inhibits haem-dependent PGRMC1 dimerization. FIG | |
| 40 46 PGRMC1 protein Carbon monoxide inhibits haem-dependent PGRMC1 dimerization. FIG | |
| 47 59 dimerization oligomeric_state Carbon monoxide inhibits haem-dependent PGRMC1 dimerization. FIG | |
| 4 33 UV-visible absorption spectra evidence (a) UV-visible absorption spectra of PGRMC1 (a.a.44–195). FIG | |
| 37 43 PGRMC1 protein (a) UV-visible absorption spectra of PGRMC1 (a.a.44–195). FIG | |
| 49 55 44–195 residue_range (a) UV-visible absorption spectra of PGRMC1 (a.a.44–195). FIG | |
| 35 46 presence of protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 51 59 oxidized protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 68 72 haem chemical Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 74 80 ferric protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 87 94 reduced protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 103 107 haem chemical Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 109 116 ferrous protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 126 133 reduced protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 142 146 haem chemical Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 152 154 CO chemical Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 160 167 ferrous protein_state Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 168 170 CO chemical Measurements were performed in the presence of the oxidized form of haem (ferric), the reduced form of haem (ferrous) and the reduced form of haem plus CO gas (ferrous+CO). FIG | |
| 21 34 haem stacking bond_interaction (b) Close-up view of haem stacking. FIG | |
| 4 33 Gel-filtration chromatography experimental_method (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 46 52 PGRMC1 protein (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 58 64 44–195 residue_range (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 66 75 wild-type protein_state (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 77 79 wt protein_state (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 89 94 Y113F mutant (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 98 103 C129S mutant (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 104 110 mutant protein_state (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 118 126 presence protein_state (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 130 140 absence of protein_state (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 141 145 haem chemical (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 147 157 dithionite chemical (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 165 167 CO chemical (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 219 225 PGRMC1 protein (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 241 245 haem chemical (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 250 252 CO chemical (c) Gel-filtration chromatography analyses of PGRMC1 (a.a.44–195) wild-type (wt) and the Y113F or C129S mutant in the presence or absence of haem, dithionite and/or CO. (d) Transition model for structural regulation of PGRMC1 in response to haem and CO. FIG | |
| 0 4 Haem chemical Haem-dependent dimerization of PGRMC1 is necessary for tumour proliferation mediated by EGFR signalling. FIG | |
| 15 27 dimerization oligomeric_state Haem-dependent dimerization of PGRMC1 is necessary for tumour proliferation mediated by EGFR signalling. FIG | |
| 31 37 PGRMC1 protein Haem-dependent dimerization of PGRMC1 is necessary for tumour proliferation mediated by EGFR signalling. FIG | |
| 88 92 EGFR protein_type Haem-dependent dimerization of PGRMC1 is necessary for tumour proliferation mediated by EGFR signalling. FIG | |
| 9 15 PGRMC1 protein (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 16 25 wild-type protein_state (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 27 29 wt protein_state (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 35 40 Y113F mutant (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 41 47 mutant protein_state (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 62 68 44–195 residue_range (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 81 84 apo protein_state (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 89 99 haem-bound protein_state (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 111 120 incubated experimental_method (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 135 139 EGFR protein_type (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 144 165 co-immunoprecipitated experimental_method (a) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo- or haem-bound form, were incubated with purified EGFR and co-immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 42 58 Western blotting experimental_method Input and bound proteins were detected by Western blotting. FIG | |
| 42 58 Western blotting experimental_method Input and bound proteins were detected by Western blotting. FIG | |
| 4 26 In vitro binding assay experimental_method (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 57 67 haem-bound protein_state (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 73 79 PGRMC1 protein (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 80 82 wt protein_state (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 88 94 44–195 residue_range (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 109 113 EGFR protein_type (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 143 148 RuCl3 chemical (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 153 158 CORM3 chemical (b) In vitro binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt (a.a.44–195) and purified EGFR with or without treatment of RuCl3 and CORM3. FIG | |
| 9 15 PGRMC1 protein (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 16 18 wt protein_state (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 22 27 Y113F mutant (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 29 40 full length protein_state (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 46 60 over-expressed experimental_method (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 81 99 immunoprecipitated experimental_method (c) FLAG-PGRMC1 wt or Y113F (full length) was over-expressed in HCT116 cells and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 0 21 Co-immunoprecipitated experimental_method Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 37 43 PGRMC1 protein Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 45 55 endogenous protein_state Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 56 62 PGRMC1 protein Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 67 71 EGFR protein_type Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 92 108 Western blotting experimental_method Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 123 129 PGRMC1 protein Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 138 142 EGFR protein_type Co-immunoprecipitated proteins (FLAG-PGRMC1, endogenous PGRMC1 and EGFR) were detected with Western blotting by using anti-PGRMC1 or anti-EGFR antibody. FIG | |
| 62 77 succinylacetone chemical (d) HCT116 cells were treated with or without 250 μmol l−1 of succinylacetone (SA) for 48 h. The intracellular haem was extracted and quantified by reverse-phase HPLC. FIG | |
| 79 81 SA chemical (d) HCT116 cells were treated with or without 250 μmol l−1 of succinylacetone (SA) for 48 h. The intracellular haem was extracted and quantified by reverse-phase HPLC. FIG | |
| 111 115 haem chemical (d) HCT116 cells were treated with or without 250 μmol l−1 of succinylacetone (SA) for 48 h. The intracellular haem was extracted and quantified by reverse-phase HPLC. FIG | |
| 148 166 reverse-phase HPLC experimental_method (d) HCT116 cells were treated with or without 250 μmol l−1 of succinylacetone (SA) for 48 h. The intracellular haem was extracted and quantified by reverse-phase HPLC. FIG | |
| 54 70 Student's t-test experimental_method of four separate experiments. **P<0.01 using unpaired Student's t-test. (e) Co-immunoprecipitation assay was performed as in (c) with or without SA treatment in HCT116 cells. FIG | |
| 76 104 Co-immunoprecipitation assay experimental_method of four separate experiments. **P<0.01 using unpaired Student's t-test. (e) Co-immunoprecipitation assay was performed as in (c) with or without SA treatment in HCT116 cells. FIG | |
| 145 147 SA chemical of four separate experiments. **P<0.01 using unpaired Student's t-test. (e) Co-immunoprecipitation assay was performed as in (c) with or without SA treatment in HCT116 cells. FIG | |
| 36 41 shRNA chemical (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 51 64 knocking down experimental_method (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 65 71 PGRMC1 protein (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 73 82 PGRMC1-KD mutant (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 102 105 EGF protein_type (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 147 151 EGFR protein_type (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 187 203 Western blotting experimental_method (f) HCT116 cells expressing control shRNA or those knocking down PGRMC1 (PGRMC1-KD) were treated with EGF or left untreated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 48 51 EGF protein_type (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 53 55 SA chemical (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 57 62 RuCl3 chemical (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 67 72 CORM3 chemical (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 109 113 EGFR protein_type (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 149 165 Western blotting experimental_method (g,h) HCT116 cells were treated with or without EGF, SA, RuCl3 and CORM3 as indicated, and components of the EGFR signaling pathway were detected by Western blotting. FIG | |
| 0 4 Haem chemical Haem-dependent dimerization of PGRMC1 accelerates tumour growth through the EGFR signaling pathway. FIG | |
| 15 27 dimerization oligomeric_state Haem-dependent dimerization of PGRMC1 accelerates tumour growth through the EGFR signaling pathway. FIG | |
| 31 37 PGRMC1 protein Haem-dependent dimerization of PGRMC1 accelerates tumour growth through the EGFR signaling pathway. FIG | |
| 76 80 EGFR protein_type Haem-dependent dimerization of PGRMC1 accelerates tumour growth through the EGFR signaling pathway. FIG | |
| 28 34 PGRMC1 protein (a) Nucleotide sequences of PGRMC1 targeted by shRNA and of the shRNA-resistant full length PGRMC1 expression vector. FIG | |
| 47 52 shRNA chemical (a) Nucleotide sequences of PGRMC1 targeted by shRNA and of the shRNA-resistant full length PGRMC1 expression vector. FIG | |
| 64 79 shRNA-resistant protein_state (a) Nucleotide sequences of PGRMC1 targeted by shRNA and of the shRNA-resistant full length PGRMC1 expression vector. FIG | |
| 80 91 full length protein_state (a) Nucleotide sequences of PGRMC1 targeted by shRNA and of the shRNA-resistant full length PGRMC1 expression vector. FIG | |
| 92 98 PGRMC1 protein (a) Nucleotide sequences of PGRMC1 targeted by shRNA and of the shRNA-resistant full length PGRMC1 expression vector. FIG | |
| 7 23 PGRMC1-knockdown mutant Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 25 34 PGRMC1-KD mutant Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 54 77 transiently transfected experimental_method Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 87 102 shRNA-resistant protein_state Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 103 120 expression vector experimental_method Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 124 133 wild-type protein_state Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 134 140 PGRMC1 protein Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 142 144 wt protein_state Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 153 158 Y113F mutant Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 159 165 mutant protein_state Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 167 172 Y113F mutant Stable PGRMC1-knockdown (PGRMC1-KD) HCT116 cells were transiently transfected with the shRNA-resistant expression vector of wild-type PGRMC1 (wt) or the Y113F mutant (Y113F). FIG | |
| 4 13 Erlotinib chemical (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 51 60 PGRMC1-KD mutant (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 70 79 PGRMC1-KD mutant (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 97 112 shRNA-resistant protein_state (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 113 119 PGRMC1 protein (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 120 122 wt protein_state (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 126 131 Y113F mutant (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 168 177 MTT assay experimental_method (b) Erlotinib was added to HCT116 (control) cells, PGRMC1-KD cells or PGRMC1-KD cells expressing shRNA-resistant PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 30 32 *P evidence of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 44 49 ANOVA experimental_method of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 55 73 Fischer's LSD test experimental_method of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 30 32 *P evidence of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 44 49 ANOVA experimental_method of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 55 73 Fischer's LSD test experimental_method of four separate experiments. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 38 47 PGRMC1-KD mutant (c) Spheroid formation in control and PGRMC1-KD HCT116 cells. FIG | |
| 54 56 *P evidence The graph represents mean±s.e. of each spheroid size. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 68 73 ANOVA experimental_method The graph represents mean±s.e. of each spheroid size. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 79 97 Fischer's LSD test experimental_method The graph represents mean±s.e. of each spheroid size. *P<0.01 using ANOVA with Fischer's LSD test. FIG | |
| 74 96 intrasplenic injection experimental_method Scale bar: 0.1 mm. (d) Tumour-bearing livers of NOG mice at 10 days after intrasplenic injection of HCT116 (control) or PGRMC1-KD cells. FIG | |
| 120 129 PGRMC1-KD mutant Scale bar: 0.1 mm. (d) Tumour-bearing livers of NOG mice at 10 days after intrasplenic injection of HCT116 (control) or PGRMC1-KD cells. FIG | |
| 28 30 *P evidence of 10 separate experiments. *P<0.05 using unpaired Student's t-test. FIG | |
| 51 67 Student's t-test experimental_method of 10 separate experiments. *P<0.05 using unpaired Student's t-test. FIG | |
| 0 4 Haem chemical Haem-dependent PGRMC1 dimerization enhances tumour chemoresistance through interaction with cytochromes P450. FIG | |
| 15 21 PGRMC1 protein Haem-dependent PGRMC1 dimerization enhances tumour chemoresistance through interaction with cytochromes P450. FIG | |
| 22 34 dimerization oligomeric_state Haem-dependent PGRMC1 dimerization enhances tumour chemoresistance through interaction with cytochromes P450. FIG | |
| 92 108 cytochromes P450 protein_type Haem-dependent PGRMC1 dimerization enhances tumour chemoresistance through interaction with cytochromes P450. FIG | |
| 11 17 PGRMC1 protein (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 18 27 wild-type protein_state (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 29 31 wt protein_state (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 37 42 Y113F mutant (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 43 49 mutant protein_state (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 64 70 44–195 residue_range (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 83 86 apo protein_state (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 90 100 haem-bound protein_state (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 112 121 incubated experimental_method (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 127 133 CYP1A2 protein (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 141 147 CYP3A4 protein (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 156 174 immunoprecipitated experimental_method (a,b) FLAG-PGRMC1 wild-type (wt) and Y113F mutant proteins (a.a.44–195), in either apo or haem-bound form, were incubated with CYP1A2 (a) or CYP3A4 (b) and immunoprecipitated with anti-FLAG antibody-conjugated beads. FIG | |
| 4 17 Binding assay experimental_method (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 48 58 haem-bound protein_state (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 64 70 PGRMC1 protein (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 71 73 wt protein_state (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 78 84 CYP1A2 protein (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 101 106 RuCl3 chemical (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 111 116 CORM3 chemical (c) Binding assay was performed as in (a) using haem-bound FLAG-PGRMC1 wt and CYP1A2 with or without RuCl3 and CORM3. FIG | |
| 30 41 doxorubicin chemical (d) Schematic illustration of doxorubicin metabolism is shown on the left. FIG | |
| 0 11 Doxorubicin chemical Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 16 25 incubated experimental_method Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 63 68 shRNA chemical Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 72 80 shPGRMC1 chemical Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 82 91 PGRMC1-KD mutant Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 102 115 doxorubicinol chemical Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 116 127 doxorubicin chemical Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 173 178 LC-MS experimental_method Doxorubicin was incubated with HCT116 cells expressing control shRNA or shPGRMC1 (PGRMC1-KD), and the doxorubicinol/doxorubicin ratios in cell pellets were determined using LC-MS. FIG | |
| 31 33 *P evidence of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 69 85 Student's t-test experimental_method of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 112 123 doxorubicin chemical of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 162 171 PGRMC1-KD mutant of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 182 191 PGRMC1-KD mutant of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 209 224 shRNA-resistant protein_state of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 225 236 full-length protein_state of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 237 243 PGRMC1 protein of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 244 246 wt protein_state of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 250 255 Y113F mutant of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 292 301 MTT assay experimental_method of four separate experiments. **P<0.01 versus control using unpaired Student's t-test. (e) Indicated amounts of doxorubicin were added to HCT116 (control) cells, PGRMC1-KD cells, or PGRMC1-KD cells expressing shRNA-resistant full-length PGRMC1 wt or Y113F, and cell viability was examined by MTT assay. FIG | |
| 40 46 PGRMC1 protein Schematic diagram for the regulation of PGRMC1 functions. FIG | |
| 0 3 Apo protein_state Apo-PGRMC1 exists as an inactive monomer. FIG | |
| 4 10 PGRMC1 protein Apo-PGRMC1 exists as an inactive monomer. FIG | |
| 24 32 inactive protein_state Apo-PGRMC1 exists as an inactive monomer. FIG | |
| 33 40 monomer oligomeric_state Apo-PGRMC1 exists as an inactive monomer. FIG | |
| 3 13 binding to protein_state On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 14 18 haem chemical On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 20 26 PGRMC1 protein On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 35 40 dimer oligomeric_state On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 49 70 stacking interactions bond_interaction On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 83 87 haem chemical On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 112 118 PGRMC1 protein On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 136 140 EGFR protein_type On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 145 161 cytochromes P450 protein_type On binding to haem, PGRMC1 forms a dimer through stacking interactions between the haem moieties, which enables PGRMC1 to interact with EGFR and cytochromes P450, leading to an enhanced proliferation and chemoresistance of cancer cells. FIG | |
| 0 2 CO chemical CO interferes with the stacking interactions of the haems and thereby inhibits PGRMC1 functions. FIG | |
| 23 44 stacking interactions bond_interaction CO interferes with the stacking interactions of the haems and thereby inhibits PGRMC1 functions. FIG | |
| 52 57 haems chemical CO interferes with the stacking interactions of the haems and thereby inhibits PGRMC1 functions. FIG | |
| 79 85 PGRMC1 protein CO interferes with the stacking interactions of the haems and thereby inhibits PGRMC1 functions. FIG | |
| 39 51 dimerization oligomeric_state PGRMC1 proteins exhibit haem-dependent dimerization in solution. TABLE | |
| 360 365 C129S mutant " Apo form Haem-bound form Mass (Da) Mass (Da) aPGRMC1 wt (a.a.44–195) ESI-MS — 17,844.14 — 36,920.19 Theoretical 17,843.65 36,918.06 Hydrodynamic radius 10−9 (m) MW (kDa) Hydrodynamic radius 10−9 (m) MW (kDa) DOSY 2.04–2.15 20 2.94–3.02 42 S20,w (S) MW (kDa) S20,w (S) MW (kDa) SV-AUC 1.9 17.6 3.1 35.5 bPGRMC1 C129S (a.a.44–195) ESI-MS — 17,827.91 — 36,887.07 Theoretical 17,827.59 36,885.6 S20,w (S) MW (kDa) S20,w (S) MW (kDa) SV-AUC 2.0 18.1 3.1 35.8 " TABLE | |
| 66 71 C129S mutant Differences in molecular weights of the wild-type (wt; a) and the C129S mutant (b) PGRMC1 proteins in the absence (apo form) or the presence of haem (haem-bound form). TABLE | |
| 32 37 C129S mutant The protein sizes of the wt and C129S PGRMC1 cytosolic domains (a.a.44–195) in the presence or absence of haem were estimated by ESI-MS, DOSY and SV-AUC. TABLE | |