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membrane protein folding

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https://www.readbyqxmd.com/read/28448574/fusion-expression-of-occludin-extracellular-loops-and-an-%C3%AE-helical-bundle-a-new-research-model-for-tight-junction
#1
Xiaojing Chi, Xia Zhao, Wei Wang, Yuqiang Niu, Min Cheng, Xiuying Liu, Sheng Cui, Wei Yang
Tight junctions (TJs) are the outermost structures of intercellular junctions and are highly specialized membrane domains involved in many important cellular processes. However, most TJ proteins are four-time transmembrane proteins and are difficult to express in their correct soluble form, which limits their functional study and therapeutic application. Human occludin (OCLN) is a major component of TJs and an essential co-receptor for hepatitis C virus (HCV) cell entry. To explore expression strategy for recombinant TJ proteins possessing integrated and functional extracellular loops, OCLN was here used as a model molecule, and several prokaryotic fusion constructs were designed by docking OCLN extracellular loops (ECLs) to HIV-1 gp41 NHR and CHR six-helical bundle (6HV1); then their biophysical features and anti-HCV activity were evaluated...
2017: PloS One
https://www.readbyqxmd.com/read/28447389/outer-membrane-vesicles-from-fibrobacter-succinogenes-s85-contain-an-array-of-carbohydrate-active-enzymes-with-versatile-polysaccharide-degrading-capacity
#2
Magnus Ø Arntzen, Anikó Várnai, Roderick I Mackie, Vincent G H Eijsink, Phillip B Pope
Fibrobacter succinogenes is an anaerobic bacterium naturally colonizing the rumen and cecum of herbivores where it utilizes an enigmatic mechanism to deconstruct cellulose into cellobiose and glucose, which serve as carbon sources for growth. Here, we illustrate that outer membrane vesicles (OMVs) released by F. succinogenes are enriched with carbohydrate-active enzymes and that intact OMVs were able to depolymerize a broad range of linear and branched hemicelluloses and pectin, despite the inability of F. succinogenes to utilize non-cellulosic (pentose) sugars for growth...
April 26, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28443122/membrane-proteomics-of-arabidopsis-glucosinolate-mutants-cyp79b2-b3-and-myb28-29
#3
Islam Mostafa, Mi-Jeong Yoo, Ning Zhu, Sisi Geng, Craig Dufresne, Maged Abou-Hashem, Maher El-Domiaty, Sixue Chen
Glucosinolates (Gls) constitute a major group of natural metabolites represented by three major classes (aliphatic, indolic and aromatic) of more than 120 chemical structures. In our previous work, soluble proteins and metabolites in Arabidopsis mutants deficient of aliphatic (myb28/29) and indolic Gls (cyp79B2B3) were analyzed. Here we focus on investigating the changes at the level of membrane proteins in these mutants. Our LC/MS-MS analyses of tandem mass tag (TMT) labeled peptides derived from the cyp79B2/B3 and myb28/29 relative to wild type resulted in the identification of 4,673 proteins, from which 2,171 are membrane proteins...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28442431/reducing-isoform-complexity-of-human-tetraspanins-by-optimized-expression-in-dictyostelium-discoideum-enables-high-throughput-functional-read-out
#4
Tineke Scheltz, Julia von Bülow, Eric Beitz
The human tetraspanin family of scaffold proteins comprises 33 isoforms. Being integral membrane proteins, they organize a so-called tetraspanin web via homomeric and heteromeric protein-protein interactions with integrins, immunoglobulins, growth factors, receptor tyrosine kinases, proteases, signaling proteins, and viral capsid proteins. Tetraspanins promote cellular effects, such as adhesion, migration, invasion, signaling, membrane fusion, protein trafficking, cancer progression, and infections. The ubiquitous expression of multiple tetraspanin isoforms and partner proteins hampers specific interaction studies...
April 22, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28435443/occurrence-of-plastidial-triacylglycerol-synthesis-and-the-potential-regulatory-role-of-agpat-in-the-model-diatom-phaeodactylum-tricornutum
#5
Srinivasan Balamurugan, Xiang Wang, Hong-Lei Wang, Chun-Jing An, Hui Li, Da-Wei Li, Wei-Dong Yang, Jie-Sheng Liu, Hong-Ye Li
BACKGROUND: Microalgae have emerged as a potential feedstock for biofuels and bioactive components. However, lack of microalgal strains with promising triacylglycerol (TAG) content and desirable fatty acid composition have hindered its commercial feasibility. Attempts on lipid overproduction by metabolic engineering remain largely challenging in microalgae. RESULTS: In this study, a microalgal 1-acyl-sn-glycerol-3-phosphate acyltransferase designated AGPAT1 was identified in the model diatom Phaeodactylum tricornutum...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28434130/characterization-of-extracellular-membrane-vesicles-of-an-antarctic-bacterium-shewanella-livingstonensis-ac10-and-their-enhanced-production-by-alteration-of-phospholipid-composition
#6
Fumiaki Yokoyama, Jun Kawamoto, Tomoya Imai, Tatsuo Kurihara
A cold-adapted bacterium, Shewanella livingstonensis Ac10, which produces eicosapentaenoic acid (EPA) as a component of its membrane phospholipids, is useful as a model to study the function of EPA and as a host for heterologous production of thermolabile proteins at low temperatures. In this study, we characterized extracellular membrane vesicles (EMVs) of this bacterium to examine the involvement of EPA in the biogenesis of EMVs and for the future application of EMVs to extracellular protein production. We found that this strain produced EMVs from the cell surface...
April 22, 2017: Extremophiles: Life Under Extreme Conditions
https://www.readbyqxmd.com/read/28433717/response-a-chronic-effects-of-pbde-47-up-regulations-of-hsp60-and-hsp70-expression-in-freshwater-bivalve-anodonta-woodiana
#7
Xichao Xia, Shipeng Xue, Xiying Wang, Qingyuan Zhang, Chuanfeng Huang, Lianghong Guo, Lunguang Yao
Heat shock proteins (HSPs) play an important role in adaption of environmental stress by protein folding, membrane translocation, degradation of misfolded proteins and other regulatory processes. Our previous study showed oxidative stress generated from polybrominated diphenyl ether-47 (PBDE-47) could cause an acute toxicity on freshwater bivalve Anodonta Woodiana, but the effect of chronic toxicity need to be elucidated. In order to further investigate the chronic effect of PBDE-47, clams A. Woodiana were randomly divided into the PBDE-47 treated group administrated with PBDE-47 at a concentration 3...
April 19, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28433053/the-envelope-proteins-of-the-bunyavirales
#8
Pablo Guardado-Calvo, Félix A Rey
The Bunyavirales Order encompasses nine families of enveloped viruses containing a single-stranded negative-sense RNA genome divided into three segments. The small (S) and large (L) segments encode proteins participating in genome replication in the infected cell cytoplasm. The middle (M) segment encodes the viral glycoproteins Gn and Gc, which are derived from a precursor polyprotein by host cell proteases. Entry studies are available only for a few viruses in the Order, and in each case they were shown to enter cells via receptor-mediated endocytosis...
2017: Advances in Virus Research
https://www.readbyqxmd.com/read/28432015/blatm-2-0-a-genetic-tool-revealing-preferred-antiparallel-interaction-of-transmembrane-helix-4-of-the-dual-topology-protein-emre
#9
Ayse Julius, Lisa Laur, Christoph Schanzenbach, Dieter Langosch
Parallel and antiparallel transmembrane helix-helix interactions support the folding and non-covalent assembly of many integral membrane proteins. While several genetic tools are currently in use to study parallel transmembrane helix-helix interactions, antiparallel associations have been difficult to determine. Here, we present a novel genetic approach, termed BLaTM 2.0, which can be used in combination with the recently presented BLaTM 1.2 to compare the efficiency of antiparallel and parallel TMD interactions in a natural membrane...
April 18, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28431525/membrane-enriched-proteome-changes-and-prion-protein-expression-during-neural-differentiation-and-in-neuroblastoma-cells
#10
J A Macedo, D Schrama, I Duarte, E Tavares, J Renaut, M E Futschik, P M Rodrigues, E P Melo
BACKGROUND: The function of the prion protein, involved in the so-called prion diseases, remains a subject of intense debate and the possibility that it works as a pleiotropic protein through the interaction with multiple membrane proteins is somehow supported by recent reports. Therefore, the use of proteomic and bioinformatics combined to uncover cellular processes occurring together with changes in the expression of the prion protein may provide further insight into the putative pleiotropic role of the prion protein...
April 22, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28431522/comparative-proteomic-analysis-of-neisseria-meningitidis-wildtype-and-dpra-null-mutant-strains-links-dna-processing-to-pilus-biogenesis
#11
Getachew Tesfaye Beyene, Shewit Kalayou, Tahira Riaz, Tone Tonjum
BACKGROUND: DNA processing chain A (DprA) is a DNA binding protein which is ubiquitous in bacteria, and is required for DNA transformation to various extents among bacterial species. However, the interaction of DprA with competence and recombination proteins is poorly understood. Therefore, the proteomes of whole Neisseria meningitidis (Nm) wildtype and dprA mutant cells were compared. Such a comparative proteomic analysis increases our understanding of the interactions of DprA with other Nm components and may elucidate its potential role beyond DNA processing in transformation...
April 21, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28430241/probing-the-germination-kinetics-of-ethanol-treated-bacillus-thuringiensis-spores
#12
Guiwen Wang, Huanjun Chen, Xiaochun Wang, Lixin Peng, Yuan Peng, Yong-Qing Li
Bacillus thuringiensis (Bt) is the most widely used microbial insecticide. To clarify the mechanism of bacterial resistance to ethanol toxicity, the present study investigated the effects of 70% (v/v) ethanol at a moderate temperature (65°C) on Bt spore germination by single-cell Raman spectroscopy and differential interference contrast microscopy. We found that over 80% of Bt spores were inviable after 30 min of treatment. Moreover, ethanol treatment affected spore germination; the time for initiation of rapid calcium dipicolinate (CaDPA) release (i...
April 20, 2017: Applied Optics
https://www.readbyqxmd.com/read/28429509/co2-permeability-and-carbonic-anhydrase-activity-of-rat-cardiomyocytes
#13
Mariela Arias-Hidalgo, Samer Al-Samir, Natalie Weber, Cornelia Geers-Knörr, Gerolf Gros, Volker Endeward
AIM: We determine here the CO2 permeability (PCO2 ) of plasma membranes of cardiomyocytes. These cells were chosen because heart possesses the highest rate of O2 -consumption/CO2 -production in the body. METHODS: Cardiomyocytes were isolated from rat hearts using the Langendorff technique. Cardiomyocyte suspensions exhibited a vitality of 2-14% and were studied by the previously described mass spectrometric (18) O-exchange-technique deriving PCO2 . We show by mass spectrometry and by carbonic anhydrase (CA)-staining that non-vital cardiomyocytes are free of CA and thus do not contribute to the mass spectrometric signal, which is determined exclusively by the fully functional vital cardiomyocytes...
April 20, 2017: Acta Physiologica
https://www.readbyqxmd.com/read/28427940/characterization-of-the-rax1-gene-encoding-a-putative-regulator-of-g-protein-signaling-in-aspergillus-fumigatus
#14
Olumuyiwa Ayokunle Igbalajobi, Jae-Hyuk Yu, Kwang-Soo Shin
The filamentous fungus Aspergillus fumigatus is the major cause of life threatening invasive aspergillosis, and its small hydrophobic asexual spores (conidia) are the major infection agent. To better understand biology of A. fumigatus, we have characterized the rax1 gene encoding a putative regulator of G protein signaling (RGS). The deletion (Δ) of rax1 results in restricted colony growth and highly reduced number of conidia in A. fumigatus. Transcript levels of the three central activators of asexual development abaA, brlA, and wetA are significantly reduced in the Δrax1 mutant...
April 17, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28425971/%C3%AF-%C3%A2-cells-feature-de-ubiquitination-of-slc-transporters-and-increased-levels-and-fluxes-of-amino-acids
#15
André Bordinassi Medina, Marcin Banaszczak, Yang Ni, Ina Aretz, David Meierhofer
Solute carrier (SLC) transporters are a diverse group of membrane transporter proteins that regulate the cellular flux and distribution of endogenous and xenobiotic compounds. Post-translational modifications (PTMs), such as ubiquitination, have recently emerged as one of the major regulatory mechanisms in protein function and localization. Previously, we showed that SLC amino acid transporters were on average 6-fold de-ubiquitinated and increased amino acid levels were detected in ρ⁰ cells (lacking mitochondrial DNA, mtDNA) compared to parental cells...
April 20, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28425077/unexpected-and-striking-effect-of-heparin-free-dialysis-on-cytokine-release
#16
Alicja Rydzewska-Rosolowska, Joanna Gozdzikiewicz-Lapinska, Jacek Borawski, Ewa Koc-Zorawska, Michal Mysliwiec, Beata Naumnik
Heparin (both unfractionated and low molecular weight) is not only a potent anticoagulant but also has many pleiotropic effects, some of which are mediated by cytokine release. We compared the effect of hemodialysis (HD) with enoxaparin as an anticoagulant and without systemic anticoagulation (heparin-grafted membrane-Evodial) on the release of monocyte chemoattractant protein 1 (MCP-1), endostatin (ES) and activin A (Act-A). Nineteen stable HD patients were dialyzed with or without heparin, and plasma levels of MCP-1, ES and Act-A were measured after such a dialysis...
April 19, 2017: International Urology and Nephrology
https://www.readbyqxmd.com/read/28423006/transcriptome-and-proteome-responses-in-rnalater-preserved-tissue-of-arabidopsis-thaliana
#17
Colin P S Kruse, Proma Basu, Darron R Luesse, Sarah E Wyatt
Tissue preservation is a minimal requirement for the success of plant RNA and protein expression studies. The standard of snap-freezing in liquid nitrogen is not always practical or possible. RNAlater, a concentrated solution of ammonium and cesium sulfates, has become a standard preservative in the absence of liquid nitrogen. Here, we demonstrate the effectiveness of RNAlater in preserving both RNA and proteins in Arabidopsis thaliana tissues for use in RNAseq and LC-MS/MS analysis of proteins. While successful in preserving plant material, a transcriptomic and proteomic response is evident...
2017: PloS One
https://www.readbyqxmd.com/read/28416748/differential-expression-of-m3-muscarinic-receptors-in-progressive-colon-neoplasia-and-metastasis
#18
Kunrong Cheng, Aaron C Shang, Cinthia B Drachenberg, Min Zhan, Jean-Pierre Raufman
M3 muscarinic receptor (M3R) activation promotes colon cancer cell proliferation, migration, and invasion in vitro. Although over-expression of CHRM3, the gene encoding M3R, is reported in primary colon cancers, expression of M3R itself has not been studied in colon neoplasia. We compared M3R expression in normal colon to colon adenomas, and primary and metastatic colon cancers. Compared to adjacent normal colon, CHRM3 expression was increased up to 128-fold in 10 of 18 consecutive surgical cancer specimens (56%) and associated with metastatic spread (P < 0...
March 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28414919/functional-characterization-of-an-n-terminally-truncated-mitochondrial-porin-expressed-in-i-neurospora-crassa-i
#19
Sabbir Rahman Shuvo, Uliana Kovaltchouk, Abdullah Zubaer, Ayush Kumar, William A T Summers, Lynda J Donald, George Hausner, Deborah A Court
Mitochondrial porin, which forms voltage-dependent anion-selective channels (VDAC) in the outer membrane, can be folded into a 19 β-stranded barrel. The N-terminus of the protein is external to the barrel, and contains α-helical structure. Targeted modifications of the N‑terminal region have been assessed in artificial membranes, leading to different models for gating <i>in vitro</i>. However, the i<i>n vivo</i> requirements for gating and the N-terminal segment of porin are less well-understood...
April 17, 2017: Canadian Journal of Microbiology
https://www.readbyqxmd.com/read/28413881/molecular-mechanism-of-stabilizing-the-helical-structure-of-huntingtin-n17-in-a-micellar-environment
#20
Leili Zhang, Hongsuk Kang, Francisco X Vázquez, Leticia Toledo-Sherman, Binquan Luan, Ruhong Zhou
Huntington's disease is a deadly neurodegenerative disease caused by the fibrilization of huntingtin (HTT) exon-1 protein mutants. Despite extensive efforts over the past decade, much remains unknown about the structures of (mutant) HTT exon-1 and their enigmatic roles in aggregation. Particularly, whether the first 17 residues in the N-terminal (HTT-N17) adopt a helical or a coiled structure remains unclear. Here, with the rigorous study of molecular dynamics simulations, we explored the most possible structures of HTT-N17 in both dodecylphosphocholine (DPC) micelles and aqueous solution, using three commonly applied force fields (OPLS-AA/L, CHARMM36, and AMBER99sb*-ILDNP) to examine the underlying molecular mechanisms and rule out potential artifacts...
April 26, 2017: Journal of Physical Chemistry. B
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