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membrane protein folding

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https://www.readbyqxmd.com/read/28944442/protein-folding-and-trna-biology
#1
REVIEW
Mónica Marín, Tamara Fernández-Calero, Ricardo Ehrlich
Polypeptides can fold into tertiary structures while they are synthesized by the ribosome. In addition to the amino acid sequence, protein folding is determined by several factors within the cell. Among others, the folding pathway of a nascent polypeptide can be affected by transient interactions with other proteins, ligands, or the ribosome, as well as by the translocation through membrane pores. Particularly, the translation machinery and the population of tRNA under different physiological or adaptive responses can dramatically affect protein folding...
September 24, 2017: Biophysical Reviews
https://www.readbyqxmd.com/read/28944418/endogenous-epitope-tagging-of-heat-shock-protein-70-isoform-hsc70-using-crispr-cas9
#2
Nitika, Andrew W Truman
Heat shock protein 70 (Hsp70) is an evolutionarily well-conserved molecular chaperone involved in several cellular processes such as folding of proteins, modulating protein-protein interactions, and transport of proteins across the membrane. Binding partners of Hsp70 (known as "clients") are identified on an individual basis as researchers discover their particular protein of interest binds to Hsp70. A full complement of Hsp70 interactors under multiple stress conditions remains to be determined. A promising approach to characterizing the Hsp70 "interactome" is the use of protein epitope tagging and then affinity purification followed by mass spectrometry (AP-MS/MS)...
September 24, 2017: Cell Stress & Chaperones
https://www.readbyqxmd.com/read/28943390/the-e-coli-s30-lysate-proteome-a-prototype-for-cell-free-protein-production
#3
Daniel Foshag, Erik Henrich, Ekkehard Hiller, Miriam Schäfer, Christian Kerger, Anke Burger-Kentischer, Irene Diaz-Moreno, Sofía M García-Mauriño, Volker Dötsch, Steffen Rupp, Frank Bernhard
Protein production using processed cell lysates is a core technology in synthetic biology and these systems are excellent to produce difficult toxins or membrane proteins. However, the composition of the central lysate of cell-free systems is still a "black box". Escherichia coli lysates are most productive for cell-free expression, yielding several mgs of protein per ml of reaction. Their preparation implies proteome fractionation, resulting in strongly biased and yet unknown lysate compositions. Many metabolic pathways are expected to be truncated or completely removed...
September 21, 2017: New Biotechnology
https://www.readbyqxmd.com/read/28943271/transmembrane-%C3%AE-barrels-evolution-folding-and-energetics
#4
REVIEW
Deepti Chaturvedi, Radhakrishnan Mahalakshmi
The biogenesis of transmembrane β-barrels (outer membrane proteins, or OMPs) is an elaborate multistep orchestration of the nascent polypeptide with translocases, barrel assembly machinery, and helper chaperone proteins. Several theories exist that describe the mechanism of chaperone-assisted OMP assembly in vivo and unassisted (spontaneous) folding in vitro. Structurally, OMPs of bacterial origin possess even-numbered strands, while mitochondrial β-barrels are even- and odd-stranded. Several underlying similarities between prokaryotic and eukaryotic β-barrels and their folding machinery are known; yet, the link in their evolutionary origin is unclear...
September 21, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28942523/overexpression-of-plastidic-maize-nadp-malate-dehydrogenase-zmnadp-mdh-in-arabidopsis-thaliana-confers-tolerance-to-salt-stress
#5
Deepika Kandoi, Sasmita Mohanty, Baishnab C Tripathy
The plastidic C4 Zea mays NADP-malate dehydrogenase (ZmNADP-MDH), responsible for catalysis of oxaloacetate to malate, was overexpressed in Arabidopsis thaliana to assess its impact on photosynthesis and tolerance to salinity stress. Different transgenic lines were produced having ~3-6-fold higher MDH protein abundance and NADP-MDH enzyme activity than vector control. The overexpressors had similar chlorophyll, carotenoid, and protein content as that of vector control. Their photosynthetic electron transport rates, carbon assimilation rate, and consequently fresh weight and dry weight were almost similar...
September 24, 2017: Protoplasma
https://www.readbyqxmd.com/read/28941661/molecular-analysis-of-human-solute-carrier-slc26-anion-transporter-disease-causing-mutations-using-3-dimensional-homology-modeling
#6
Chloe Rapp, Xiaoyun Bai, Reinhart A F Reithmeier
The availability of the first crystal structure of a bacterial member (SLC26Dg) of the solute carrier SLC26 family of anion transporters has allowed us to create 3-dimensional models of all 10 human members (SLC26A1-A11, A10 being a pseudogene) of these membrane proteins using the Phyre2 bioinformatic tool. The homology modeling predicted that the SLC26 human proteins, like the SLC26Dg template, all consist of 14 transmembrane segments (TM) arranged in a 7+7 inverted topology with the amino-termini of two half-helices (TM3 and 10) facing each other in the centre of the protein to create the anion-binding site, linked to a C-terminal cytosolic sulfate transporter anti-sigma factor antagonist (STAS) domain...
September 20, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28939593/boosted-coupling-of-atp-hydrolysis-to-substrate-transport-upon-cooperative-estradiol-17-%C3%AE-d-glucuronide-binding-in-a-drosophila-atp-binding-cassette-type-c-transporter
#7
Agnes Karasik, Kaitlyn Victoria Ledwitch, Tamás Arányi, András Váradi, Arthur Roberts, Flóra Szeri
ATP binding cassette type-C (ABCC) transporters move molecules across cell membranes upon hydrolysis of ATP; however, their coupling of ATP hydrolysis to substrate transport remains elusive. Drosophila multidrug resistance-associated protein (DMRP) is the functional ortholog of human long ABCC transporters, with similar substrate and inhibitor specificity, but higher activity. Exploiting its high activity, we kinetically dissected the catalytic mechanism of DMRP by using E2-d-glucuronide (E2G), the physiologic substrate of human ABCC...
September 22, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28939366/membrane-induced-organization-and-dynamics-of-the-n-terminal-domain-of-chemokine-receptor-cxcr1-insights-from-atomistic-simulations
#8
Shalmali Kharche, Manali Joshi, Durba Sengupta, Amitabha Chattopadhyay
The CXC chemokine receptor 1 (CXCR1) is an important member of the G protein-coupled receptor (GPCR) family in which the extracellular N-terminal domain has been implicated in ligand binding and selectivity. The structure of this domain has not yet been elucidated due to its inherent dynamics, but experimental evidence points toward membrane-dependent organization and dynamics. To gain molecular insight into the interaction of the N-terminal domain with the membrane bilayer, we performed a series of microsecond time scale atomistic simulations of the N-terminal domain of CXCR1 in the presence and absence of POPC bilayers...
September 19, 2017: Chemistry and Physics of Lipids
https://www.readbyqxmd.com/read/28933630/going-native-direct-high-throughput-screening-of-secreted-full-length-igg-antibodies-against-cell-membrane-proteins
#9
Yongliang Fang, Thach H Chu, Margaret E Ackerman, Karl E Griswold
Gel microdroplet - fluorescence activated cell sorting (GMD-FACS) is an innovative high throughput screening platform for recombinant protein libraries, and we show here that GMD-FACS can overcome many of the limitations associated with conventional screening methods for antibody libraries. For example, phage and cell surface display benefit from exceptionally high throughput, but generally require high quality, soluble antigen target and necessitate the use of anchored antibody fragments. In contrast, the GMD-FACS assay can screen for soluble, secreted, full-length IgGs at rates of several thousand clones per second, and the technique enables direct screening against membrane protein targets in their native cellular context...
September 21, 2017: MAbs
https://www.readbyqxmd.com/read/28932871/crystal-structures-of-eukaryote-glycosyltransferases-reveal-biologically-relevant-enzyme-homooligomers
#10
REVIEW
Deborah Harrus, Sakari Kellokumpu, Tuomo Glumoff
Glycosyltransferases (GTases) transfer sugar moieties to proteins, lipids or existing glycan or polysaccharide molecules. GTases form an important group of enzymes in the Golgi, where the synthesis and modification of glycoproteins and glycolipids take place. Golgi GTases are almost invariably type II integral membrane proteins, with the C-terminal globular catalytic domain residing in the Golgi lumen. The enzymes themselves are divided into 103 families based on their sequence homology. There is an abundance of published crystal structures of GTase catalytic domains deposited in the Protein Data Bank (PDB)...
September 20, 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28931852/improvement-of-a-fermentation-process-for-the-production-of-two-pfama1-dico-based-malaria-vaccine-candidates-in-pichia-pastoris
#11
Robin Kastilan, Alexander Boes, Holger Spiegel, Nadja Voepel, Ivana Chudobová, Stephan Hellwig, Johannes Felix Buyel, Andreas Reimann, Rainer Fischer
Pichia pastoris is a simple and powerful expression platform that has the ability to produce a wide variety of recombinant proteins, ranging from simple peptides to complex membrane proteins. A well-established fermentation strategy is available comprising three main phases: a batch phase, followed by a glycerol fed-batch phase that increases cell density, and finally an induction phase for product expression using methanol as the inducer. We previously used this three-phase strategy at the 15-L scale to express three different AMA1-DiCo-based malaria vaccine candidates to develop a vaccine cocktail...
September 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28931397/pseudomonas-stutzeri-as-an-alternative-host-for-membrane-proteins
#12
Manuel Sommer, Hao Xie, Hartmut Michel
BACKGROUND: Studies on membrane proteins are often hampered by insufficient yields of the protein of interest. Several prokaryotic hosts have been tested for their applicability as production platform but still Escherichia coli by far is the one most commonly used. Nevertheless, it has been demonstrated that in some cases hosts other than E. coli are more appropriate for certain target proteins. RESULTS: Here we have developed an expression system for the heterologous production of membrane proteins using a single plasmid-based approach...
September 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28931075/excitability-is-increased-in-hippocampal-ca1-pyramidal-cells-of-fmr1-knockout-mice
#13
M Angeles Luque, Pablo Beltran-Matas, M Carmen Marin, Blas Torres, Luis Herrero
Fragile X syndrome (FXS) is caused by a failure of neuronal cells to express the gene encoding the fragile mental retardation protein (FMRP). Clinical features of the syndrome include intellectual disability, learning impairment, hyperactivity, seizures and anxiety. Fmr1 knockout (KO) mice do not express FMRP and, as a result, reproduce some FXS behavioral abnormalities. While intrinsic and synaptic properties of excitatory cells in various part of the brain have been studied in Fmr1 KO mice, a thorough analysis of action potential characteristics and input-output function of CA1 pyramidal cells in this model is lacking...
2017: PloS One
https://www.readbyqxmd.com/read/28930630/assessing-the-structure-and-stability-of-transmembrane-oligomeric-intermediates-of-an-%C3%AE-helical-toxin
#14
Rajat Desikan, Prabal K K Maiti, K Ganapathy Ayappa
Protein membrane interactions play an important role in our understanding of diverse phenomena ranging from membrane assisted protein aggregation to oligomerization and folding. Pore forming toxins (PFTs) form the primary vehicle for infection by several strains of bacteria. These proteins expressed in a water soluble form (monomers) bind to the target membrane (protomer) to form a transmembrane pore complex through a process of oligomerization. Based on the structure of the transmembrane domains, PFTs are broadly classified into β or α toxins...
September 20, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28929861/ulipristal-acetate-and-extracellular-matrix-production-in-human-leiomyomas-in-vivo-a-laboratory-analysis-of-a-randomized-placebo-controlled-trial
#15
Jeris Cox, Minnie Malik, Joy Britten, Terrence Lewis, William H Catherino
In a prior randomized controlled study, patients treated with ulipristal acetate (UPA) or placebo for 3 months had a decrease in leiomyoma size. A total of 10 patients' tissue samples (5 placebo and 5 treated with 10 mg/d UPA) that underwent hysterectomy and tissue preservation were identified from this study. Quantitative real-time reverse transcriptase polymerase chain reaction and Western blotting were used to assess fold gene and protein expression of extracellular membrane (ECM) proteins: collagen 1A (COL1A), fibronectin (FN1), and versican (VCAN) of the samples...
January 1, 2017: Reproductive Sciences
https://www.readbyqxmd.com/read/28927344/structural-elucidation-of-transmembrane-domain-zero-tmd0-of-ecdl-a-multidrug-resistance-associated-protein-mrp-family-of-atp-binding-cassette-transporter-protein-revealed-by-atomistic-simulation
#16
Krishnendu Bera, Priyanka Rani, Gaurav Kishor, Shikha Agarwal, Antresh Kumar, Durg Vijay Singh
ATP-Binding cassette (ABC) transporters play an extensive role in the translocation of diverse sets of biologically important molecules across membrane. EchnocandinB (antifungal) and EcdL protein of Aspergillus rugulosus are encoded by the same cluster of genes. Co-expression of EcdL and echinocandinB reflects tightly linked biological functions. EcdL belongs to Multidrug Resistance associated Protein (MRP) subfamily of ABC transporters with an extra transmembrane domain zero (TMD0). Complete structure of MRP subfamily comprising of TMD0 domain, at atomic resolution is not known...
September 20, 2017: Journal of Biomolecular Structure & Dynamics
https://www.readbyqxmd.com/read/28924263/refinement-of-ectopic-protein-expression-through-the-gal4-uas-system-in-bombyx-mori-application-to-behavioral-and-developmental-studies
#17
Chiho Hara, Koudai Morishita, Seika Takayanagi-Kiya, Akihisa Mikami, Keiro Uchino, Takeshi Sakurai, Ryohei Kanzaki, Hideki Sezutsu, Masafumi Iwami, Taketoshi Kiya
Silkmoth, Bombyx mori, is one of the important model insects in which transgenic techniques and the GAL4/UAS system are applicable. However, due to cytotoxicity and low transactivation activity of GAL4, effectiveness of the GAL4/UAS system and its application in B. mori are still limited. In the present study, we refined the previously reported UAS vector by exploiting transcriptional and translational enhancers, and achieved 200-fold enhancement of reporter GFP fluorescence in the GAL4/UAS system. Enhanced protein expression of membrane-targeted GFP and calcium indicator protein (GCaMP5G) drastically improved visualization of fine neurite structures and neural activity, respectively...
September 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28924037/intrinsically-disordered-sequences-enable-modulation-of-protein-phase-separation-through-distributed-tyrosine-motifs
#18
Yuan Lin, Simon L Currie, Michael Keith Rosen
Liquid-liquid phase separation (LLPS) is thought to contribute to establishment of many biomolecular condensates, eukaryotic cell structures that concentrate diverse macromolecules but lack a bounding membrane. RNA granules control RNA metabolism and comprise a large class of condensates that are enriched in RNA-binding proteins and RNA molecules. Many RNA granule proteins are composed of both modular domains and intrinsically disordered regions (IDRs) having low amino acid sequence complexity. Phase separation of these molecules likely plays an important role in the generation and stability of RNA granules...
September 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28923515/integrative-functions-of-the-mitochondrial-contact-site-and-cristae-organizing-system
#19
REVIEW
Stefan Schorr, Martin van der Laan
Mitochondria are complex double-membrane-bound organelles of eukaryotic cells that function as energy-converting powerhouses, metabolic factories and signaling centers. The outer membrane controls the exchange of material and information with other cellular compartments. The inner membrane provides an extended, highly folded surface for selective transport and energy-coupling reactions. It can be divided into an inner boundary membrane and tubular or lamellar cristae membranes that accommodate the oxidative phosphorylation units...
September 15, 2017: Seminars in Cell & Developmental Biology
https://www.readbyqxmd.com/read/28921397/a-binding-block-ion-selective-mechanism-revealed-by-a-na-k-selective-channel
#20
Jie Yu, Bing Zhang, Yixiao Zhang, Cong-Qiao Xu, Wei Zhuo, Jingpeng Ge, Jun Li, Ning Gao, Yang Li, Maojun Yang
Mechanosensitive (MS) channels are extensively studied membrane protein for maintaining intracellular homeostasis through translocating solutes and ions across the membrane, but its mechanisms of channel gating and ion selectivity are largely unknown. Here, we identified the YnaI channel as the Na(+)/K(+) cation-selective MS channel and solved its structure at 3.8 Å by cryo-EM single-particle method. YnaI exhibits low conductance among the family of MS channels in E. coli, and shares a similar overall heptamer structure fold with previously studied MscS channels...
September 18, 2017: Protein & Cell
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