bluuebunny/crossref_metadata_2025 huggingface dataset:
BCL11B enhances TCR/CD28-triggered NF-κB activation through up-regulation of Cot kinase gene expression in T-lymphocytes
Authors: Cismasiu Valeriu B. , Duque Javier , Paskaleva Elena , Califano Danielle , Ghanta Sailaja , Young Howard A. , Avram Dorina
Published: October 2008

BCL11B is a transcriptional regulator with an important role in T-cell development and leukaemogenesis. We demonstrated recently that BCL11B controls expression from the IL (interleukin)-2 promoter through direct binding to the US1 (upstream site 1). In the present study, we provide evidence that BCL11B also participates in the activation of IL-2 gene expression by enhancing NF-κB (nuclear factor κB) activity in the context of TCR (T-cell receptor)/CD28-triggered T-cell activation. Enhanced NF-κB activation is not a consequence of BCL11B binding to the NF-κB response elements or association with the NF-κB–DNA complexes, but rather the result of higher translocation of NF-κB to the nucleus caused by enhanced degradation of IκB (inhibitor of NF-κB). The enhanced IκB degradation in cells with increased levels of BCL11B was specific for T-cells activated through the TCR, but not for cells activated through TNFα (tumour necrosis factor α) or UV light, and was caused by increased activity of IκB kinase, as indicated by its increase in phosphorylation. As BCL11B is a transcription factor, we investigated whether the expression of genes upstream of IκB kinase in the TCR/CD28 signalling pathway was affected by increased BCL11B expression, and found that Cot (cancer Osaka thyroid oncogene) kinase mRNA levels were elevated. Cot kinase is known to promote enhanced IκB kinase activity, which results in the phosphorylation and degradation of IκB and activation of NF-κB. The implied involvement of Cot kinase in BCL11B-mediated NF-κB activation in response to TCR activation is supported by the fact that a Cot kinase dominant-negative mutant or Cot kinase siRNA (small interfering RNA) knockdown blocked BCL11B-mediated NF-κB activation. In support of our observations, in the present study we report that BCL11B enhances the expression of several other NF-κB target genes, in addition to IL-2. In addition, we provide evidence that BCL11B associates with intron 2 of the Cot kinase gene to regulate its expression.
Phosphorylated and non-phosphorylated forms of catechol O-methyltransferase in rat liver, brain and other tissues
Authors: Øverbye Anders , Seglen Per O.
Published: October 2008

Seven different forms of the enzyme COMT (catechol O-methyltransferase) were found in isolated rat hepatocytes by two-dimensional gel electrophoresis and immunoblotting: five small variants (S-COMT) and two large variants (L-COMT). The identities of these COMT forms were verified by tryptic fingerprinting using MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS, and by amino acid sequencing using ESI–IT–MS/MS (electrospray ionization with ion-trap tandem MS). Analysis of tissue distributions showed that the S-COMT forms were highly expressed in liver and kidney, whereas L-COMT was expressed more strongly in other tissues. Both of the L-COMT forms were found in all of the tissues examined except the heart, which expressed only the most acidic form, and the kidney, which expressed only the most basic form. Subcellular fractionation revealed the presence of both S-COMT and L-COMT in soluble, as well as sedimentable, fractions, suggesting that they should be classified by size rather than (as previously) by localization. Several of the S-COMT forms were N-acetylated, and the two most acidic forms were found to be phosphorylated at Ser260. One of the latter was unique to liver cells; the other was also found in kidney, brain and thymus. Among the non-phosphorylated S-COMT forms, one was ubiquitous, one was found in testis and liver, and a third was found in liver, kidney and thymus. No other phosphorylated sites were found, suggesting that the pI differences distinguishing between the various COMT forms are due to some as yet unidentified structural modification(s).
Characterization of cytochrome bo3 activity in a native-like surface-tethered membrane
Authors: Weiss Sophie A. , Bushby Richard J. , Evans Stephen D. , Henderson Peter J. F. , Jeuken Lars J. C.
Published: October 2008

We have developed a simple native-like surface-tethered membrane system to investigate the activity of cbo3 (cytochrome bo3), a terminal oxidase in Escherichia coli. The tethered membranes consist of E. coli inner-membrane extracts mixed with additional E. coli lipids containing various amounts of the cbo3 substrate UQ-10 (ubiquinol-10). Tethered membranes are formed by self-assembly from vesicles on to gold electrodes functionalized with cholesterol derivatives. cbo3 activity was monitored using CV (cyclic voltammetry) with electron transfer to cbo3 mediated by UQ-10. The apparent Km for oxygen with this system is 1.1±0.4 μM, in good agreement with values reported in the literature for whole-cell experiments and for purified cbo3. Increasing the concentration of lipophilic UQ-10 in the membrane leads to an increase in cbo3 activity. The activity of cbo3 with long-chain ubiquinones appears to be different from previous reports using short-chain substrate analogues such as UQ-1 in that typical Michaelis–Menten kinetics are not observed using UQ-10. This native-like membrane model thus provides new insights into the interaction of transmembrane enzymes with hydrophobic substrates which contrasts with studies using hydrophilic UQ analogues.
Yeast chitin synthase 2 activity is modulated by proteolysis and phosphorylation
Authors: Martínez-Rucobo Fuensanta W. , Eckhardt-Strelau Luise , Terwisscha Van Scheltinga Anke C.
Published: October 2008

Saccharomyces cerevisiae Chs2 (chitin synthase 2) synthesizes the primary septum after mitosis is completed. It is essential for proper cell separation and is expected to be highly regulated. We have expressed Chs2 and a mutant lacking the N-terminal region in Pichia pastoris in an active form at high levels. Both constructs show a pH and cation dependence similar to the wild-type enzyme, as well as increased activity after trypsin treatment. Using further biochemical analysis, we have identified two mechanisms of chitin synthase regulation. First, it is hyperactivated by a soluble yeast protease. This protease is expressed during exponential growth phase, when budding cells require Chs2 activity. Secondly, LC-MS/MS (liquid chromatography tandem MS) experiments on purified Chs2 identify 12 phosphorylation sites, all in the N-terminal domain. Four of them show the perfect sequence motif for phosphorylation by the cyclin-dependent kinase Cdk1. As we also show that phosphorylation of the N-terminal domain is important for Chs2 stability, these sites might play an important role in the cell cycle-dependent degradation of the enzyme, and thus in cell division.
Phagocytosis: a repertoire of receptors and Ca2+ as a key second messenger
Authors: Melendez Alirio J. , Tay Hwee Kee
Published: October 2008

Receptor-mediated phagocytosis is a complex process that mediates the internalization, by a cell, of other cells and large particles; this is an important physiological event not only in mammals, but in a wide diversity of organisms. Of simple unicellular organisms that use phagocytosis to extract nutrients, to complex metazoans in which phagocytosis is essential for the innate defence system, as a first line of defence against invading pathogens, as well as for the clearance of damaged, dying or dead cells. Evolution has armed multicellular organisms with a range of receptors expressed on many cells that serve as the molecular basis to bring about phagocytosis, regardless of the organism or the specific physiological event concerned. Key to all phagocytic processes is the finely controlled rearrangement of the actin cytoskeleton, in which Ca2+ signals play a major role. Ca2+ is involved in cytoskeletal changes by affecting the actions of a number of contractile proteins, as well as being a cofactor for the activation of a number of intracellular signalling molecules, which are known to play important roles during the initiation, progression and resolution of the phagocytic process. In mammals, the requirement of Ca2+ for the initial steps in phagocytosis, and the subsequent phagosome maturation, can be quite different depending on the type of cell and on the type of receptor that is driving phagocytosis. In this review we discuss the different receptors that mediate professional and non-professional phagocytosis, and discuss the role of Ca2+ in the different steps of this complex process.
Cln6 mutants associated with neuronal ceroid lipofuscinosis are degraded in a proteasome-dependent manner
Authors: Oresic Kristina , Mueller Britta , Tortorella Domenico
Published: October 2008

NCLs (neuronal ceroid lipofuscinoses), a group of inherited neurodegenerative lysosomal storage diseases that predominantly affect children, are the result of autosomal recessive mutations within one of the nine cln genes. The wild-type cln gene products are composed of membrane and soluble proteins that localize to the lysosome or the ER (endoplasmic reticulum). However, the destiny of the Cln variants has not been fully characterized. To explore a possible link between ER quality control and processing of Cln mutants, we investigated the fate of two NCL-related Cln6 mutants found in patient samples (Cln6G123D and Cln6M241T) in neuronal-derived human cells. The point mutations are predicted to be in the putative transmembrane domains and most probably generate misfolded membrane proteins that are subjected to ER quality control. Consistent with this paradigm, both mutants underwent rapid proteasome-mediated degradation and complexed with components of the ER extraction apparatus, Derlin-1 and p97. In addition, knockdown of SEL1L [sel-1 suppressor of lin-12-like (Caenorhabditis elegans)], a member of an E3 ubiquitin ligase complex involved in ER protein extraction, rescued significant amounts of Cln6G123D and Cln6M241T polypeptides. The results implicate ER quality control in the instability of the Cln variants that probably contributes to the development of NCL.
Versatile role of the yeast ubiquitin ligase Rsp5p in intracellular trafficking
Authors: Belgareh-Touzé Naima , Léon Sébastien , Erpapazoglou Zoi , Stawiecka-Mirota Marta , Urban-Grimal Danièle , Haguenauer-Tsapis Rosine
Published: October 2008

The ubiquitin ligase (E3) Rsp5p is the only member of the Nedd (neural-precursor-cell-expressed, developmentally down-regulated) 4 family of E3s present in yeast. Rsp5p has several proteasome-independent functions in membrane protein trafficking, including a role in the ubiquitination of most plasma membrane proteins, leading to their endocytosis. Rsp5p is also required for the ubiquitination of endosomal proteins, leading to their sorting to the internal vesicles of MVBs (multivesicular bodies). Rsp5p catalyses the attachment of non-conventional ubiquitin chains, linked through ubiquitin Lys-63, to some endocytic and MVB cargoes. This modification appears to be required for efficient sorting, possibly because these chains have a greater affinity for the ubiquitin-binding domains present within endocytic or MVB sorting complexes. The mechanisms involved in the recognition of plasma membrane and MVB substrates by Rsp5p remain unclear. A subset of Rsp5/Nedd4 substrates have a ‘PY motif’ and are recognized directly by the WW (Trp-Trp) domains of Rsp5p. Most Rsp5p substrates do not carry PY motifs, but some may depend on PY-containing proteins for their ubiquitination by Rsp5p, consistent with the latter's acting as specificity factors or adaptors. As in other ubiquitin-conjugating systems, these adaptors are also Rsp5p substrates and undergo ubiquitin-dependent trafficking. In the present review, we discuss recent examples illustrating the role of Rsp5p in membrane protein trafficking and providing new insights into the regulation of this E3 by adaptor proteins.
Ubiquitin ligase E6-AP and its role in human disease
Authors: Matentzoglu Konstantin , Scheffner Martin
Published: October 2008

The ubiquitin ligase E6-AP (E6-associated protein) represents a prime example for the notion that deregulated modification of proteins with ubiquitin contributes to the development of human disease: loss of E6-AP function by mutation is responsible for the development of AS (Angelman syndrome), a neurological disorder, and unscheduled activation of E6-AP by complex formation with the E6 oncoprotein of HPVs (human papillomaviruses) contributes to cervical carcinogenesis. However, while there is a considerable amount of data concerning the oncogenic properties of the E6–E6-AP complex, only little is known about the function(s) of E6-AP in neurons. This is mainly due to the fact that although some E6-AP substrates have been identified, it is at present unclear whether deregulated modification/degradation of these proteins is involved in the pathogenesis of AS. Similarly, the cellular pathways involving E6-AP remain enigmatic. To obtain insights into the physiological functions of E6-AP, we are currently employing several strategies, including quantitative affinity proteomics and RNA interference approaches. The results obtained will eventually allow the introduction of E6-AP into functional protein networks and so reveal potential targets for molecular approaches in the treatment of E6-AP-associated diseases.
Novel substrates and functions for the ubiquitin-like molecule NEDD8
Authors: Xirodimas Dimitris P.
Published: October 2008

Genetic experiments have established an important role for the ubiquitin-like molecule NEDD8 (neural-precursor-cell-expressed developmentally down-regulated 8) in the regulation of cell growth, viability and development. It is therefore essential to identify the molecular targets for the pathway. Until recently, the cullin family of proteins was characterized as the only substrates for NEDDylation. However, through either direct biological approaches or the use of proteomics, it is now evident that the NEDD8 proteome is more diverse than thought previously. The present review describes the biological significance of NEDDylation for the novel identified substrates and the emerging evidence for the co-operation between the ubiquitin and NEDD8 pathways to control protein function.
How autophagy is related to programmed cell death during the development of the nervous system
Authors: Boya Patricia , Mellén María Angeles , de la Rosa Enrique J.
Published: October 2008

Programmed cell death, together with proliferation and differentiation, is an essential process during the development of the nervous system. During neurogenesis, neurons and glia are generated in large numbers and, subsequently, they die in a process that depends on trophic signalling that refines the cytoarchitecture and connectivity of the nervous system. In addition, programmed cell death also affects proliferating neuroepithelial cells and recently differentiated neuroblasts. Autophagy is a lysosomal degradative pathway that allows the recycling of cell constituents, and seems to be able to play a dual role. It may serve to protect the cell by preventing the accumulation of deleterious products and organelles and supplying energy and amino acids. On the other hand, it has been considered a type of cell death. The role of autophagy during development is little characterized. The retina provides an excellent model system to study autophagy in the context of neural development, and to establish its relationship with proliferation, differentiation and cell death. In the present review, we summarize recent findings showing that autophagy contributes to the development of the nervous system by providing energy for cell corpse removal after physiological cell death, a process associated with retinal neurogenesis.
Regulation of ER-associated degradation via p97/VCP-interacting motif
Authors: Ballar Petek , Fang Shengyun
Published: October 2008

p97/VCP (valosin-containing protein) is a cytosolic AAA (ATPase associated with various cellular activities) essential for retrotranslocation of misfolded proteins during ERAD [ER (endoplasmic reticulum)-associated degradation]. gp78, an ERAD ubiquitin ligase, is one of the p97/VCP recruitment proteins localized to the ER membrane. A newly identified VIM (p97/VCP-interacting motif) in gp78 has brought about novel insights into mechanisms of ERAD, such as the presence of a p97/VCP-dependent but Ufd1-independent retrotranslocation during gp78-mediated ERAD. Additionally, SVIP (small p97/VCP-interacting protein), which contains a VIM in its N-terminal region, negatively regulates ERAD by uncoupling p97/VCP and Derlin1 from gp78. Thus SVIP may protect cells from damage by extravagant ERAD.
Fos family protein degradation by the proteasome
Authors: Gomard Tiphanie , Jariel-Encontre Isabelle , Basbous Jihane , Bossis Guillaume , Mocquet-Torcy Gabriel , Piechaczyk Marc
Published: October 2008

c-Fos proto-oncoprotein defines a family of closely related transcription factors (Fos proteins) also comprising Fra-1, Fra-2, FosB and ΔFosB, the latter two proteins being generated by alternative splicing. Through the regulation of many genes, most of them still unidentified, they regulate major functions from the cell level up to the whole organism. Thus they are involved in the control of proliferation, differentiation and apoptosis, as well as in the control of responses to stresses, and they play important roles in organogenesis, immune responses and control of cognitive functions, among others. Fos proteins are intrinsically unstable. We have studied how two of them, c-Fos and Fra-1, are degraded. Departing from the classical scenario where unstable key cell regulators are hydrolysed by the proteasome after polyubiquitination, we showed that the bulk of c-Fos and Fra-1 can be broken down independently of any prior ubiquitination. Certain conserved structural domains suggest that similar mechanisms may also apply to Fra-2 and FosB. Computer search indicates that certain motifs shared by the Fos proteins and putatively responsible for instability are found in no other protein, suggesting the existence of degradation mechanisms specific for this protein family. Under particular signalling conditions, others have shown that a part of cytoplasmic c-Fos requires ubiquitination for fast turnover. This poses the question of the multiplicity of degradation pathways that apply to proteins depending on their intracellular localization.
Lipases in the pancreatic β-cell: implications for insulin secretion
Authors: Fex Malin , Mulder Hindrik
Published: October 2008

Lipids have been implicated in β-cell stimulus–secretion coupling. In such a role, lipases in β-cells would be required to generate lipid coupling factors. We have shown previously that glucose stimulates lipolysis in rodent islets. In addition, lipolysis and diacylglycerol lipase activity in islets are abolished by orlistat, an irreversible lipase inhibitor with a broad specificity for substrates. Moreover, orlistat dose-dependently inhibits glucose- and forskolin-stimulated insulin secretion, while leaving glucose oxidation and the rise in the ATP/ADP ratio intact. In an effort to identify β-cell lipase(s), we found that HSL (hormone-sensitive lipase), the rate-limiting enzyme for acylglycerol hydrolysis in adipocytes, is expressed in rodent β-cells. To resolve the role of this lipase, we have created global and β-cell-specific knockout mice. Although our line of global HSL-knockout mice is moderately glucose-intolerant owing to reduced peripheral insulin sensitivity and exhibits normal islet metabolism and insulin secretion, other HSL-knockout lines have displayed impaired insulin secretion under certain conditions. In contrast, β-cell-specific HSL-knockout mice, which are less prone to genetic redundancy, are hyperglycaemic, presumably caused by a perturbation of first-phase insulin secretion. Thus studies by us and others demonstrate that lipases, such as HSL, play a regulatory role in β-cell stimulus–secretion coupling.
The cytoprotective actions of long-chain mono-unsaturated fatty acids in pancreatic β-cells
Authors: Morgan Noel G. , Dhayal Shalinee , Diakogiannaki Eleftheria , Welters Hannah J.
Published: October 2008

Chronic exposure of pancreatic β-cells to long-chain fatty acids can cause loss of secretory function and enhanced apoptosis by a process of ‘lipotoxicity’, which may be a contributory factor to the rising incidence of Type 2 diabetes in humans. However, when incubated in vitro, β-cells respond differentially to long-chain saturated and mono-unsaturated fatty acids, suggesting that these molecules may regulate cell functionality by different mechanisms. In particular, it is clear that, whereas saturated fatty acids [e.g. palmitate (C16:0)] exert detrimental effects on β-cells, the equivalent mono-unsaturated species [e.g. palmitoleate (C16:1)] are well tolerated. Indeed, mono-unsaturated species are potently cytoprotective. The present review explores the differential effects of these various fatty acids on β-cell viability and considers the possible mechanisms involved in cytoprotection by mono-unsaturates.
An update on lipotoxic endoplasmic reticulum stress in pancreatic β-cells
Authors: Cnop Miriam , Igoillo-Esteve Mariana , Cunha Daniel A. , Ladrière Laurence , Eizirik Décio L.
Published: October 2008

The UPR (unfolded protein response) or ER (endoplasmic reticulum) stress response was first described 20 years ago. The field of ER stress has expanded tremendously since, moving from basic biology in yeast to human neurodegenerative, inflammatory, cardiovascular and neoplastic diseases. The ER stress response has also been implicated in diabetes development, affecting both insulin production by pancreatic β-cells and insulin sensitivity in peripheral tissues. In the present mini-review, we focus on recent progress in the field of ER stress in pancreatic β-cells. Recent advances in the understanding of lipotoxic ER stress and β-cell recovery from ER stress are discussed.
NOX family NADPH oxidases in liver and in pancreatic islets: a role in the metabolic syndrome and diabetes?
Authors: Guichard Cécile , Moreau Richard , Pessayre Dominique , Epperson Terry Kay , Krause Karl-Heinz
Published: October 2008

The incidence of obesity and non-esterified (‘free’) fatty acid-associated metabolic disorders such as the metabolic syndrome and diabetes is increasing dramatically in most countries. Although the pathogenesis of these metabolic disorders is complex, there is emerging evidence that ROS (reactive oxygen species) are critically involved in the aberrant signalling and tissue damage observed in this context. Indeed, it is now widely accepted that ROS not only play an important role in physiology, but also contribute to cell and tissue dysfunction. Inappropriate ROS generation may contribute to tissue dysfunction in two ways: (i) dysregulation of redox-sensitive signalling pathways, and (ii) oxidative damage to biological structures (DNA, proteins, lipids, etc.). An important source of ROS is the NOX family of NADPH oxidases. Several NOX isoforms are expressed in the liver and pancreatic β-cells. There is now evidence that inappropriate activation of NOX enzymes may damage the liver and pancreatic β-cells. In the context of the metabolic syndrome, the emerging epidemic of non-alcoholic steatohepatitis is thought to be NOX/ROS-dependent and of particular medical relevance. NOX/ROS-dependent β-cell damage is thought to be involved in glucolipotoxicity and thereby leads to progression from the metabolic syndrome to Type 2 diabetes. Thus understanding the role of NOX enzymes in liver and β-cell damage should lead to an increased understanding of pathomechanisms in the metabolic syndrome and diabetes and may identify useful targets for novel therapeutic strategies.
Molecular mechanisms of proteinuria in diabetes
Authors: Gnudi Luigi
Published: October 2008

The epidemic of Type 2 diabetes, and the parallel rising incidence of end-stage renal disease, is progressively increasing worldwide. Kidney disease is one of the major chronic microvascular complications of diabetes, and both metabolic and haemodynamic perturbations participate in its development and progression towards end-stage renal disease. Hypertension and poor metabolic control seem to interact in causing the relentless decline in renal function seen in diabetic patients. Both high circulating glucose levels and increased glomerular capillary pressure act in conjunction in stimulating the different cellular pathways leading to kidney disease. It has been suggested that mechanical forces at the glomerular level may aggravate the metabolic insult by stimulating excessive cellular glucose uptake by up-regulating the facilitative GLUT-1 (glucose transporter-1). We propose the existence of a self-maintaining cellular mechanism whereby a haemodynamic stimulus on glomerular cells induces the up-regulation of GLUT-1, an event followed by greater glucose uptake and activation of intracellular metabolic pathways, resulting in excess TGF-β1 (transforming growth factor-β1) production. TGF-β1, one of the major prosclerotic cytokines in diabetic kidney disease, maintains the up-regulation of GLUT-1, perpetuating a series of cellular events that result, as their ultimate effect, in increased extracellular matrix synthesis and altered permeability of the glomerular filtration barrier. Mechanical and metabolic coupling could represent an important mechanism of injury in the diabetic kidney.
The long-chain fatty acid receptor, GPR40, and glucolipotoxicity: investigations using GPR40-knockout mice
Authors: Brownlie Ruth , Mayers Rachel M. , Pierce Jackie A. , Marley Anna E. , Smith David M.
Published: October 2008

GPR40 (G-protein-coupled receptor 40) has been shown to be a physiologically relevant receptor for long-chain fatty acids. It is a family A G-protein-coupled receptor highly expressed in the β-cell where it increases insulin secretion by signalling via Gq and phospholipase C. Fatty acids are well known to mediate both acute stimulatory effects and chronic detrimental effects on the β-cell. GPR40-transgenic and GPR40−/− animals have been important tools in studies of the metabolic effects of GPR40. In the present article, we review the literature on transgenic GPR40 models and present some of our own studies on the effects of a high-fat diet on the metabolic phenotype of GPR40−/− mice. GPR40 ligands represent interesting novel therapies for Type 2 diabetes but it is presently unclear whether agonists or antagonists represent the best therapeutic approach.
Saturated and unsaturated (including arachidonic acid) non-esterified fatty acid modulation of insulin secretion from pancreatic β-cells
Authors: Keane Deirdre , Newsholme Philip
Published: October 2008

Both stimulatory and detrimental effects of NEFAs (non-esterified fatty acids) on pancreatic β-cells have been recognized. Acute exposure of the pancreatic β-cell to high glucose concentrations and/or saturated NEFAs results in a substantial increase in insulin release, whereas chronic exposure results in desensitization and suppression of secretion followed by induction of apoptosis. Some unsaturated NEFAs also promote insulin release acutely, but they are less toxic to β-cells during chronic exposure and can even exert positive protective effects. In the present review, we focus on exogenous and endogenous effects of NEFAs, including the polyunsaturated fatty acid, arachidonic acid (or its metabolites generated from cyclo-oxygenase activity), on β-cell metabolism, and have explored the outcomes with respect to β-cell insulin secretion.
Nucleotide-induced conformational changes in the Escherichia coli NADH:ubiquinone oxidoreductase (complex I)
Authors: Pohl Thomas , Schneider Daniel , Hielscher Ruth , Stolpe Stefan , Dörner Katerina , Kohlstädt Markus , Böttcher Bettina , Hellwig Petra , Friedrich Thorsten
Published: October 2008

The energy-converting NADH:ubiquinone oxidoreductase, also known as respiratory complex I, couples the transfer of electrons from NADH to ubiquinone with the translocation of protons across the membrane. Electron microscopy revealed the two-part structure of the complex consisting of a peripheral and a membrane arm. The peripheral arm contains all known cofactors and the NADH-binding site, whereas the membrane arm has to be involved in proton translocation. Owing to this, a conformation-linked mechanism for redox-driven proton translocation is discussed. By means of electron microscopy, we show that both arms of the Escherichia coli complex I are widened after the addition of NADH but not of NADPH. NADH-induced conformational changes were also detected in solution: ATR-FTIR (attenuated total reflection Fourier-transform infrared) of the soluble NADH dehydrogenase fragment of the complex indicates protein re-arrangements induced by the addition of NADH. EPR spectroscopy of surface mutants of the complex containing a covalently bound spin label at distinct positions demonstrates NADH-dependent conformational changes in both arms of the complex.
Electroneutral and electrogenic catalysis by dihaem-containing succinate:quinone oxidoreductases
Authors: Lancaster C. Roy D. , Herzog Elena , Juhnke Hanno D. , Madej M. Gregor , Müller Florian G. , Paul Rajsekhar , Schleidt Philipp G.
Published: October 2008

Membrane protein complexes can support both the generation and utilization of a transmembrane electrochemical proton potential (Δp), either by supporting transmembrane electron transfer coupled to protolytic reactions on opposite sides of the membrane or by supporting transmembrane proton transfer. Regarding the first mechanism, this has been unequivocally demonstrated to be operational for Δp-dependent catalysis of succinate oxidation by quinone in the case of the dihaem-containing SQR (succinate:menaquinone reductase) from the Gram-positive bacterium Bacillus licheniformis. This is physiologically relevant in that it allows the transmembrane Δp to drive the endergonic oxidation of succinate by menaquinone by the dihaem-containing SQR of Gram-positive bacteria. In the case of a related but different respiratory membrane protein complex, the dihaem-containing QFR (quinol:fumarate reductase) of the ϵ-proteobacterium Wolinella succinogenes, evidence has been obtained indicating that both mechanisms are combined, so as to facilitate transmembrane electron transfer by proton transfer via a both novel and essential compensatory transmembrane proton transfer pathway (‘E-pathway’). This is necessary because, although the reduction of fumarate by menaquinol is exergonic, it is obviously not exergonic enough to support the generation of a Δp. This compensatory E-pathway appears to be required by all dihaem-containing QFR enzymes and the conservation of the essential acidic residue on transmembrane helix V (Glu-C180 in W. succinogenes QFR) is a useful key for the sequence-based discrimination of these QFR enzymes from the dihaem-containing SQR enzymes.
55Mn-ENDOR of the S2-state multiline signal of Photosystem II from Thermosynechococcus elongatus
Authors: Pudollek Susanne , Lendzian Friedhelm , Bittl Robert
Published: October 2008

Pulse EPR and 55Mn-ENDOR (electron nuclear double resonance) experiments at 34 GHz (Q-band) were performed on the S2 state of the OEC (oxygen-evolving complex) in PSII (Photosystem II) from Thermosynechococcus elongatus. The ENDOR spectra are discussed with respect to earlier data obtained from X- (9 GHz) and Q-band experiments on PSII preparations from spinach with and without the addition of methanol.
Quinone-reactive proteins devoid of haem b form widespread membrane-bound electron transport modules in bacterial respiration
Authors: Simon Jörg , Kern Melanie
Published: October 2008

Many quinone-reactive enzyme complexes that are part of membrane-integral eukaryotic or prokaryotic respiratory electron transport chains contain one or more haem b molecules embedded in the membrane. In recent years, various novel proteins have emerged that are devoid of haem b but are thought to fulfil a similar function in bacterial anaerobic respiratory systems. These proteins are encoded by genes organized in various genomic arrangements and are thought to form widespread membrane-bound quinone-reactive electron transport modules that exchange electrons with redox partner proteins located at the outer side of the cytoplasmic membrane. Prototypic representatives are the multihaem c-type cytochromes NapC, NrfH and TorC (NapC/NrfH family), the putative iron–sulfur protein NapH and representatives of the NrfD/PsrC family. Members of these protein families vary in the number of their predicted transmembrane segments and, consequently, diverse quinone-binding sites are expected. Only a few of these enzymes have been isolated and characterized biochemically and high-resolution structures are limited. This mini-review briefly summarizes predicted and experimentally demonstrated properties of the proteins in question and discusses their role in electron transport and bioenergetics of anaerobic respiration.
Dicarbonyls linked to damage in the powerhouse: glycation of mitochondrial proteins and oxidative stress
Authors: Rabbani Naila , Thornalley Paul J.
Published: October 2008

Protection of mitochondrial proteins from glycation by endogenous dicarbonyl compounds, methylglyoxal and glyoxal, was found recently to prevent increased formation of reactive oxygen species and oxidative and nitrosative damage to the proteome during aging and produce life extension in the nematode Caenorhabditis elegans. This suggests that dicarbonyl glycation damage to the mitochondrial proteome may be a preceding event to mitochondrial dysfunction leading to oxidative stress. Future research will address the functional charges in mitochondrial proteins that are the targets for dicarbonyl glycation.
Cellular factors important for the de novo formation of yeast prions
Authors: Tuite Mick , Stojanovski Klement , Ness Frederique , Merritt Gloria , Koloteva-Levine Nadejda
Published: October 2008

Prions represent an unusual structural form of a protein that is ‘infectious’. In mammals, prions are associated with fatal neurodegenerative diseases such as CJD (Creutzfeldt–Jakob disease), while in fungi they act as novel epigenetic regulators of phenotype. Even though most of the human prion diseases arise spontaneously, we still know remarkably little about how infectious prions form de novo. The [PSI+] prion of the yeast Saccharomyces cerevisiae provides a highly tractable model in which to explore the underlying mechanism of de novo prion formation, in particular identifying key cis- and trans-acting factors. Most significantly, the de novo formation of [PSI+] requires the presence of a second prion called [PIN+], which is typically the prion form of Rnq1p, a protein rich in glutamine and aspartic acid residues. The molecular mechanism by which the [PIN+] prion facilitates de novo [PSI+] formation is not fully established, but most probably involves some form of cross-seeding. A number of other cellular factors, in particular chaperones of the Hsp70 (heat-shock protein 70) family, are known to modify the frequency of de novo prion formation in yeast.
The spindle pole body plays a key role in controlling mitotic commitment in the fission yeast Schizosaccharomyces pombe
Authors: Hagan Iain M.
Published: October 2008

Commitment to mitosis is regulated by a conserved protein kinase complex called MPF (mitosis-promoting factor). MPF activation triggers a positive-feedback loop that further promotes the activity of its activating phosphatase Cdc25 and is assumed to down-regulate the MPF-inhibitory kinase Wee1. Four protein kinases contribute to this amplification loop: MPF itself, Polo kinase, MAPK (mitogen-activated protein kinase) and Greatwall kinase. The fission yeast SPB (spindle pole body) component Cut12 plays a critical role in modulating mitotic commitment. In this review, I discuss the relationship between Cut12 and the fission yeast Polo kinase Plo1 in mitotic control. These results indicate that commitment to mitosis is co-ordinated by control networks on the spindle pole. I then describe how the Cut12/Plo1 control network links growth control signalling from TOR (target of rapamycin) and MAPK networks to the activation of MPF to regulate the timing of cell division.
Failure to up-regulate VEGF165b in maternal plasma is a first trimester predictive marker for pre-eclampsia
Authors: Bills Victoria L. , Varet Julia , Millar Ann , Harper Steven J. , Soothill Peter W. , Bates David O.
Published: October 2008

Pre-eclampsia is a pregnancy-related condition characterized by hypertension, proteinuria and endothelial dysfunction. VEGF165b, formed by alternative splicing of VEGF (vascular endothelial growth factor) pre-mRNA, inhibits VEGF165-mediated vasodilation and angiogenesis, but has not been quantified in pregnancy. ELISAs were used to measure means±S.E.M. plasma VEGF165b, sEng (soluble endoglin) and sFlt-1 (soluble fms-like tyrosine kinase-1). At 12 weeks of gestation, the plasma VEGF165b concentration was significantly up-regulated in plasma from women who maintained normal blood pressure throughout their pregnancy (normotensive group, 4.90±1.6 ng/ml; P<0.01, as determined using a Mann-Whitney U test) compared with non-pregnant women (0.40±0.22 ng/ml). In contrast, in patients who later developed pre-eclampsia, VEGF165b levels were lower than in the normotensive group (0.467±0.209 ng/ml), but were no greater than non-pregnant women. At term, plasma VEGF165b concentrations were greater than normal in both pre-eclamptic (3.75±2.24 ng/ml) and normotensive (10.58 ng/ml±3.74 ng/ml; P>0.1 compared with pre-eclampsia) pregnancies. Patients with a lower than median plasma VEGF165b at 12 weeks had elevated sFlt-1 and sEng pre-delivery. Concentrations of sFlt-1 (1.20±0.07 and 1.27±0.18 ng/ml) and sEng (4.4±0.18 and 4.1±0.5 ng/ml) were similar at 12 weeks of gestation in the normotensive and pre-eclamptic groups respectively. Plasma VEGF165b levels were elevated in pregnancy, but this increase is delayed in women that subsequently develop pre-eclampsia. In conclusion, low VEGF165b may therefore be a clinically useful first trimester plasma marker for increased risk of pre-eclampsia.
Lanthanide 4f-level location in lanthanide doped and cerium-lanthanide codoped NaLaF4 by photo- and thermoluminescence
Authors: Krumpel A. H. , van der Kolk E. , Zeelenberg D. , Bos A. J. J. , Krämer K. W. , Dorenbos P.
Published: October 2008

Photo- and thermoluminescence (TL) spectra of NaLaF4:Ln3+ (Ln=Ce,Pr,Nd,Sm,Eu,Gd,Tb,Dy,Ho,Er,Tm) and NaLaF4:Ce3+, Ln3+ (Ln=Nd,Sm,Ho,Er,Tm) are presented and used together with the empirical Dorenbos model in order to establish the 4f energy level positions of all tri- and divalent lanthanide ions doped in NaLaF4. The information will be presented in the form of an energy level diagram. It is shown that in addition to this diagram only two assumptions, viz., the presence of two host related electron traps and the presence of Vk-centers, are necessary for explaining the lanthanide-specific TL glow curves of both Ln mono- and Ce–Ln codoped NaLaF4.
Electronic structure of the indium tin oxide/nanocrystalline anatase (TiO2)/ruthenium-dye interfaces in dye-sensitized solar cells
Authors: Lyon J. E. , Rayan M. K. , Beerbom M. M. , Schlaf R.
Published: October 2008

The electronic structure of two interfaces commonly found in dye-sensitized photovoltaic cells based on nanocrystalline anatase TiO2 (“Grätzel cells”) was investigated using photoemission spectroscopy (PES). X-ray photoemission spectroscopy (XPS) and ultraviolet photoemission spectroscopy (UPS) measurements were carried out on the indium tin oxide (ITO)/TiO2 and the TiO2/cis-bis(isothiocyanato)bis(2,2′-bipyridyl-4,4′-dicarboxylato)-ruthenium(II)bis-tetrabutylammonium dye (“N719” or “Ruthenium 535-bisTBA”) interfaces. Both contacts were investigated using a multistep deposition procedure where the entire structure was prepared in vacuum using electrospray deposition. In between deposition steps the surface was characterized with XPS and UPS resulting in a series of spectra, allowing the determination of the orbital and band lineup at the interfaces. The results of these efforts confirm previous PES measurements on TiO2/dye contacts prepared under ambient conditions, suggesting that ambient contamination might not have significant influence on the electronic structure at the dye/TiO2 interface. The results also demonstrate that there may be a significant barrier for electron injection at the sputtered ITO/TiO2 interface and that this interface should be viewed as a semiconductor heterojunction rather than as metal-semiconductor (Schottky) contact.
Enhanced spontaneous emission in a photonic-crystal light-emitting diode
Authors: Francardi M. , Balet L. , Gerardino A. , Chauvin N. , Bitauld D. , Li L. H. , Alloing B. , Fiore A.
Published: October 2008

We report direct evidence of enhanced spontaneous emission in a photonic-crystal (PhC) light-emitting diode. The device consists of p-i-n heterojunction embedded in a suspended membrane, comprising a layer of self-assembled quantum dots. Current is injected laterally from the periphery to the center of the PhC. A well-isolated emission peak at 1.3μm from the PhC cavity mode is observed, and the enhancement of the spontaneous emission rate is clearly evidenced by time-resolved electroluminescence measurements, showing that our diode switches off in a time shorter than the bulk radiative and nonradiative lifetimes.
Spontaneous alignment of self-assembled ABC triblock terpolymers for large-area nanolithography
Authors: Kubo T. , Wang R. F. , Olson D. A. , Rodwogin M. , Hillmyer M. A. , Leighton C.
Published: October 2008

A general method for preparation of large-area inorganic nanostructure arrays based on the use of core-shell cylinder forming ABC triblock terpolymers is presented. The essential perpendicular cylinder alignment is achieved by the use of a low surface energy B block (polyisoprene), which drives spontaneous perpendicular alignment of cylindrical C domains (polylactide) in a matrix of A (polystyrene), eliminating the lengthy anneal steps, external field alignment procedures, or specialized surface preparations that could severely restrict potential applications.
The anatomy of the Gunn laser
Authors: Chung S. , Balkan N.
Published: October 2008

A monopolar GaAs Fabry–Pérot cavity laser based on the Gunn effect is studied both experimentally and theoretically. The light emission occurs via the band-to-band recombination of impact-ionized excess carriers in the propagating space-charge (Gunn) domains. Electroluminescence spectrum from the cleaved end-facet emission of devices with Ga1−xAlxAs (x=0.32) waveguides shows clearly a preferential mode at a wavelength around 840 nm at T=95 K. The threshold laser gain is assessed by using an impact ionization coefficient resulting from excess carriers inside the high-field domain.
Landau collision operators and general moment equations for an electron-ion plasma
Authors: Ji Jeong-Young , Held Eric D.
Published: October 2008

The general moment equations for an electron-ion plasma are established. The distribution functions for electrons and ions are expanded in terms of orthogonal polynomials of random velocity variables in contrast to the total velocity variables [J.-Y. Ji and E. D. Held, Phys. Plasmas 13, 102103 (2006)]. The moments of the streaming part of the kinetic equation are explicitly written with simple formulas. A simple version of the exact linearized Coulomb collision integrals is presented for like species. The electron-ion and ion-electron operators that conserve momentum and energy are also calculated with a small mass-ratio approximation. It is shown in the relaxation theory that the Lorentz operator, as a replacement of the like-species operator, is acceptable only for the high-order harmonic moments.
2
mm
waveband saturation transfer electron paramagnetic resonance of conducting polymers
Authors: Krinichnyi V. I.
Published: October 2008

The 2mm waveband (140GHz) saturation transfer electron paramagnetic resonance (ST-EPR) spectroscopy has been employed to characterize the very slow microsecond to millisecond librational macromolecular dynamics of a wide range of conducting polymers. It is possible at this waveband to determine separately spin relaxation and dynamics affecting ST-EPR spectra. Higher microwave frequency provides substantial increases sensitivity of the method to the anisotropic macromolecular motion in conducting polymers and broadens the interval of correlation times up to 1–80ms, thereby extending the slow-motion limit for ST-EPR by two orders of magnitude compared with convenient wavebands EPR.
Lamb wave attenuation in a rough plate. I. Analytical and experimental results in an anisotropic plate
Authors: Potel Catherine , Leduc Damien , Morvan Bruno , Depollier Claude , Hladky-Hennion Anne-Christine , Izbicki Jean-Louis , Pareige Pascal , Bruneau Michel
Published: October 2008

The characterization of bounded roughened surfaces before applying adhesive joint, in order to detect poor cohesive and adhesive properties, remains difficult. Earlier studies based on analysis of surface wave (Rayleigh waves or Scholte waves) are not really adapted to the characterization of such surfaces. Guided acoustic waves, i.e., Lamb waves, turn out to be the best adapted kind of waves to characterize this roughness when plates are bounded together. It is the aim of this paper to provide analytical and experimental approaches to analyze the behavior of Lamb waves propagating inside plates with a rough surface (small perturbations). First, experimental results of the attenuation effects are given on roughened glass plates. Second, the attenuation factor of the Lamb wave in an anisotropic rough solid plate is calculated through a complex analytical model of the dispersion equation which accounts for the effect of the power spectrum density of the rough profile (including the effect of the statistical roughness parameters).
Folding of polyglutamine chains
Authors: Chopra Manan , Reddy Allam S. , Abbott N. L. , de Pablo J. J.
Published: October 2008

Long polyglutamine chains have been associated with a number of neurodegenerative diseases. These include Huntington’s disease, where expanded polyglutamine (PolyQ) sequences longer than 36 residues are correlated with the onset of symptoms. In this paper we study the folding pathway of a 54-residue PolyQ chain into a β-helical structure. Transition path sampling Monte Carlo simulations are used to generate unbiased reactive pathways between unfolded configurations and the folded β-helical structure of the polyglutamine chain. The folding process is examined in both explicit water and an implicit solvent. Both models reveal that the formation of a few critical contacts is necessary and sufficient for the molecule to fold. Once the primary contacts are formed, the fate of the protein is sealed and it is largely committed to fold. We find that, consistent with emerging hypotheses about PolyQ aggregation, a stable β-helical structure could serve as the nucleus for subsequent polymerization of amyloid fibrils. Our results indicate that PolyQ sequences shorter than 36 residues cannot form that nucleus, and it is also shown that specific mutations inferred from an analysis of the simulated folding pathway exacerbate its stability.