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Membrane protein crystal structure

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https://www.readbyqxmd.com/read/28077872/structural-basis-for-nutrient-acquisition-by-dominant-members-of-the-human-gut-microbiota
#1
Amy J Glenwright, Karunakar R Pothula, Satya P Bhamidimarri, Dror S Chorev, Arnaud Baslé, Susan J Firbank, Hongjun Zheng, Carol V Robinson, Mathias Winterhalter, Ulrich Kleinekathöfer, David N Bolam, Bert van den Berg
The human large intestine is populated by a high density of microorganisms, collectively termed the colonic microbiota, which has an important role in human health and nutrition. The survival of microbiota members from the dominant Gram-negative phylum Bacteroidetes depends on their ability to degrade dietary glycans that cannot be metabolized by the host. The genes encoding proteins involved in the degradation of specific glycans are organized into co-regulated polysaccharide utilization loci, with the archetypal locus sus (for starch utilisation system) encoding seven proteins, SusA-SusG...
January 11, 2017: Nature
https://www.readbyqxmd.com/read/28076802/peldor-spectroscopy-reveals-two-defined-states-of-a-sialic-acid-trap-transporter-sbp-in-solution
#2
Janin Glaenzer, Martin F Peter, Gavin H Thomas, Gregor Hagelueken
The tripartite ATP-independent periplasmic (TRAP) transporters are a widespread class of membrane transporters in bacteria and archaea. Typical substrates for TRAP transporters are organic acids including the sialic acid N-acetylneuraminic acid. The substrate binding proteins (SBP) of TRAP transporters are the best studied component and are responsible for initial high-affinity substrate binding. To better understand the dynamics of the ligand binding process, pulsed electron-electron double resonance (PELDOR, also known as DEER) spectroscopy was applied to study the conformational changes in the N-acetylneuraminic acid-specific SBP VcSiaP...
January 10, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28069938/crystal-structure-and-mechanistic-basis-of-a-functional-homolog-of-the-antigen-transporter-tap
#3
Anne Nöll, Christoph Thomas, Valentina Herbring, Tina Zollmann, Katja Barth, Ahmad Reza Mehdipour, Thomas M Tomasiak, Stefan Brüchert, Benesh Joseph, Rupert Abele, Vincent Oliéric, Meitian Wang, Kay Diederichs, Gerhard Hummer, Robert M Stroud, Klaas M Pos, Robert Tampé
ABC transporters form one of the largest protein superfamilies in all domains of life, catalyzing the movement of diverse substrates across membranes. In this key position, ABC transporters can mediate multidrug resistance in cancer therapy and their dysfunction is linked to various diseases. Here, we describe the 2.7-Å X-ray structure of heterodimeric Thermus thermophilus multidrug resistance proteins A and B (TmrAB), which not only shares structural homology with the antigen translocation complex TAP, but is also able to restore antigen processing in human TAP-deficient cells...
January 9, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28067914/structure-of-the-immature-zika-virus-at-9-%C3%A3-resolution
#4
Vidya Mangala Prasad, Andrew S Miller, Thomas Klose, Devika Sirohi, Geeta Buda, Wen Jiang, Richard J Kuhn, Michael G Rossmann
The current Zika virus (ZIKV) epidemic is characterized by severe pathogenicity in both children and adults. Sequence changes in ZIKV since its first isolation are apparent when pre-epidemic strains are compared with those causing the current epidemic. However, the residues that are responsible for ZIKV pathogenicity are largely unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of the immature ZIKV at 9-Å resolution. The cryo-EM map was fitted with the crystal structures of the precursor membrane and envelope glycoproteins and was shown to be similar to the structures of other known immature flaviviruses...
January 9, 2017: Nature Structural & Molecular Biology
https://www.readbyqxmd.com/read/28065508/kobuviral-non-structural-3a-proteins-act-as-molecular-harnesses-to-hijack-the-host-acbd3-protein
#5
Martin Klima, Dominika Chalupska, Bartosz Różycki, Jana Humpolickova, Lenka Rezabkova, Jan Silhan, Adriana Baumlova, Anna Dubankova, Evzen Boura
Picornaviruses are small positive-sense single-stranded RNA viruses that include many important human pathogens. Within the host cell, they replicate at specific replication sites called replication organelles. To create this membrane platform, they hijack several host factors including the acyl-CoA-binding domain-containing protein-3 (ACBD3). Here, we present a structural characterization of the molecular complexes formed by the non-structural 3A proteins from two species of the Kobuvirus genus of the Picornaviridae family and the 3A-binding domain of the host ACBD3 protein...
December 21, 2016: Structure
https://www.readbyqxmd.com/read/28065269/methods-for-structural-and-functional-analyses-of-intramembrane-prenyltransferases-in-the-ubia-superfamily
#6
Y Yang, N Ke, S Liu, W Li
The UbiA superfamily is a group of intramembrane prenyltransferases that generate lipophilic compounds essential in biological membranes. These compounds, which include various quinones, hemes, chlorophylls, and vitamin E, participate in electron transport and function as antioxidants, as well as acting as structural lipids of microbial cell walls and membranes. Prenyltransferases producing these compounds are involved in important physiological processes and human diseases. These UbiA superfamily members differ significantly in their enzymatic activities and substrate selectivities...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28056928/new-ligation-independent-cloning-vectors-for-expression-of-recombinant-proteins-with-a-self-cleaving-cpd-6xhis-tag
#7
Marco Biancucci, Jazel S Dolores, Jennifer Wong, Sarah Grimshaw, Wayne F Anderson, Karla J F Satchell, Keehwan Kwon
BACKGROUND: Recombinant protein purification is a crucial step for biochemistry and structural biology fields. Rapid robust purification methods utilize various peptide or protein tags fused to the target protein for affinity purification using corresponding matrices and to enhance solubility. However, affinity/solubility-tags often need to be removed in order to conduct functional and structural studies, adding complexities to purification protocols. RESULTS: In this work, the Vibrio cholerae MARTX toxin Cysteine Protease Domain (CPD) was inserted in a ligation-independent cloning (LIC) vector to create a C-terminal 6xHis-tagged inducible autoprocessing enzyme tag, called "the CPD-tag"...
January 5, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28052259/conformational-freedom-of-the-lrp6-ectodomain-is-regulated-by-n-glycosylation-and-the-binding-of-the-wnt-antagonist-dkk1
#8
Kyoko Matoba, Emiko Mihara, Keiko Tamura-Kawakami, Naoyuki Miyazaki, Shintaro Maeda, Hidenori Hirai, Samuel Thompson, Kenji Iwasaki, Junichi Takagi
LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules...
January 3, 2017: Cell Reports
https://www.readbyqxmd.com/read/28045456/redox-potential-as-a-master-variable-controlling-pathways-of-metal-reduction-by-geobacter-sulfurreducens
#9
Caleb E Levar, Colleen L Hoffman, Aubrey J Dunshee, Brandy M Toner, Daniel R Bond
Geobacter sulfurreducens uses at least two different pathways to transport electrons out of the inner membrane quinone pool before reducing acceptors beyond the outer membrane. When growing on electrodes poised at oxidizing potentials, the CbcL-dependent pathway operates at or below redox potentials of -0.10 V vs the standard hydrogen electrode, whereas the ImcH-dependent pathway operates only above this value. Here, we provide evidence that G. sulfurreducens also requires different electron transfer proteins for reduction of a wide range of Fe(III)- and Mn(IV)-(oxyhydr)oxides, and must transition from a high- to low-potential pathway during reduction of commonly studied soluble and insoluble metal electron acceptors...
January 3, 2017: ISME Journal
https://www.readbyqxmd.com/read/28045020/structural-characterisation-of-the-fatty-acid-biosynthesis-enzyme-fabf-from-the-pathogen-listeria-monocytogenes
#10
Tatiana P Soares da Costa, Jeffrey D Nanson, Jade K Forwood
Development of new antimicrobial agents is required against the causative agent for listeriosis, Listeria monocytogenes, as the number of drug resistant strains continues to increase. A promising target is the β-ketoacyl-acyl carrier protein synthase FabF, which participates in the catalysis of fatty acid synthesis and elongation, and is required for the production of phospholipid membranes, lipoproteins, and lipopolysaccharides. In this study, we report the 1.35 Å crystal structure of FabF from L. monocytogenes, providing an excellent platform for the rational design of novel inhibitors...
January 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28042595/expression-purification-and-crystallization-of-the-herpesvirus-nuclear-egress-complex-nec
#11
Janna M Bigalke, Ekaterina E Heldwein
The protocol describes the production and crystallization of the soluble form of the nuclear egress complex (NEC) from Herpes simplex virus 1 and Pseudorabies virus. The NEC is a heterodimer that consists of conserved proteins UL31 and UL34. NEC oligomerization deforms the inner nuclear membrane around the capsid in infected cells, thereby mediating capsid budding into the perinuclear space during nuclear egress. We have successfully developed a protocol for large-scale preparation of highly pure NEC from two different viruses in a prokaryotic expression system, which enabled us to crystallize these viral protein complexes and determine their structures...
July 20, 2016: Bio-protocol
https://www.readbyqxmd.com/read/28034717/crystal-structure-of-a-phospholipase-a2-from-bothrops-asper-venom-insights-into-a-new-putative-myotoxic-cluster
#12
Guilherme H M Salvador, Juliana I Dos Santos, Bruno Lomonte, Marcos R M Fontes
Snake venoms from the Viperidae and Elapidae families often have several phospholipases A2 (PLA2s), which may display different functions despite having a similar structural scaffold. These proteins are considered an important target for the development of drugs against local myotoxic damage because they are not efficiently neutralized by conventional serum therapy. PLA2s from these venoms are generally divided into two classes: (i) catalytic PLA2s (or Asp49-PLA2s) and (ii) non-catalytic PLA2-like toxins (or Lys49-PLA2s)...
December 26, 2016: Biochimie
https://www.readbyqxmd.com/read/28030602/structural-and-biochemical-characterization-of-chlamydia-trachomatis-dsba-reveals-a-cysteine-rich-and-weakly-oxidising-oxidoreductase
#13
Signe Christensen, Morten K Grøftehauge, Karl Byriel, Wilhelmina M Huston, Emily Furlong, Begoña Heras, Jennifer L Martin, Róisín M McMahon
The Gram negative bacteria Chlamydia trachomatis is an obligate intracellular human pathogen that can cause pelvic inflammatory disease, infertility and blinding trachoma. C. trachomatis encodes a homolog of the dithiol oxidoreductase DsbA. Bacterial DsbA proteins introduce disulfide bonds to folding proteins providing structural bracing for secreted virulence factors, consequently these proteins are potential targets for antimicrobial drugs. Despite sharing functional and structural characteristics, the DsbA enzymes studied to date vary widely in their redox character...
2016: PloS One
https://www.readbyqxmd.com/read/28025687/characterisation-of-the%C3%A2-daacs%C3%A2-family-escherichia-coli-glutamate-aspartate-proton-symporter-gltp-using-computational-chemical-biochemical-and-biophysical-methods
#14
Moazur Rahman, Fouzia Ismat, Li Jiao, Jocelyn M Baldwin, David J Sharples, Stephen A Baldwin, Simon G Patching
Escherichia coli glutamate/aspartate-proton symporter GltP is a member of the Dicarboxylate/Amino Acid:Cation Symporter family of secondary active transport proteins. A range of computational, chemical, biochemical and biophysical methods characterised evolutionary relationships, structural features, substrate binding affinities and transport kinetics of wild-type and mutant forms of GltP. Sequence alignments and phylogenetic analysis revealed close homologies of GltP with human glutamate transporters involved in neurotransmission, neutral amino acid transporters and with the archaeal aspartate transporter GltPh...
December 26, 2016: Journal of Membrane Biology
https://www.readbyqxmd.com/read/28018966/crystal-structures-of-mdfa-complexed-with-acetylcholine-and-inhibitor-reserpine
#15
Ming Liu, Jie Heng, Yuan Gao, Xianping Wang
The DHA12 family of transporters contains a number of prokaryotic and eukaryote membrane proteins. Some of these proteins share conserved sites intrinsic to substrate recognition, structural stabilization and conformational changes. For this study, we chose the MdfA transporter as a model DHA12 protein to study some general characteristics of the vesicular neurotransmitter transporters (VNTs), which all belong to the DHA12 family. Two crystal structures were produced for E. coli MdfA, one in complex with acetylcholine and the other with potential reserpine, which are substrate and inhibitor of VNTs, respectively...
2016: Biophysics Reports
https://www.readbyqxmd.com/read/28011271/structural-mechanism-underlying-regulation-of-human-efhd2-swiprosin-1-actin-bundling-activity-by-ser183-phosphorylation
#16
Kyoung Ryoung Park, Jun Yop An, Jung Youn Kang, Jung-Gyu Lee, Youngjin Lee, Sang A Mun, Chang-Duk Jun, Woo Keun Song, Soo Hyun Eom
EF-hand domain-containing protein D2/Swiprosin-1 (EFhd2) is an actin-binding protein mainly expressed in the central nervous and the immune systems of mammals. Intracellular events linked to EFhd2, such as membrane protrusion formation, cell adhesion, and BCR signaling, are triggered by the association of EFhd2 and F-actin. We previously reported that Ca(2+) enhances the F-actin-bundling ability of EFhd2 through maintaining a rigid parallel EFhd2-homodimer structure. It was also reported that the F-actin-bundling ability of EFhd2 is regulated by a phosphorylation-dependent mechanism...
December 21, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28003465/structural-basis-of-influenza-virus-fusion-inhibition-by-the-antiviral-drug-arbidol
#17
Rameshwar U Kadam, Ian A Wilson
The broad-spectrum antiviral drug Arbidol shows efficacy against influenza viruses by targeting the hemagglutinin (HA) fusion machinery. However, the structural basis of the mechanism underlying fusion inhibition by Arbidol has remained obscure, thereby hindering its further development as a specific and optimized influenza therapeutic. We determined crystal structures of Arbidol in complex with influenza virus HA from pandemic 1968 H3N2 and recent 2013 H7N9 viruses. Arbidol binds in a hydrophobic cavity in the HA trimer stem at the interface between two protomers...
December 21, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28002022/interface-analysis-of-small-gtp-binding-protein-complexes-suggests-preferred-membrane-orientations
#18
Ingrid R Vetter
Crystal structures of small GTP binding protein complexes with their effectors and regulators reveal that one particularly flat side of the G domain that contains helix α4 and the C-terminal helix α5 is practically devoid of contacts. Although this observation seems trivial since the main binding targets are the switch I and II regions opposite of this side, the fact that all interacting proteins, even the largest ones, seem to avoid occupying this area (except for Ran, that does not localize to membranes) is quite striking...
December 21, 2016: Biological Chemistry
https://www.readbyqxmd.com/read/27999184/chiral-twist-drives-raft-formation-and-organization-in-membranes-composed-of-rod-like-particles
#19
Louis Kang, Tom C Lubensky
Lipid rafts are hypothesized to facilitate protein interaction, tension regulation, and trafficking in biological membranes, but the mechanisms responsible for their formation and maintenance are not clear. Insights into many other condensed matter phenomena have come from colloidal systems, whose micron-scale particles mimic basic properties of atoms and molecules but permit dynamic visualization with single-particle resolution. Recently, experiments showed that bidisperse mixtures of filamentous viruses can self-assemble into colloidal monolayers with thermodynamically stable rafts exhibiting chiral structure and repulsive interactions...
January 3, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27995422/entry-of-severe-fever-with-thrombocytopenia-syndrome-virus
#20
REVIEW
Fei Yuan, Aihua Zheng
Severe fever with thrombocytopenia syndrome virus (SFTSV) is a globe-shaped virus covered by a dense icosahedral array of glycoproteins Gn/Gc that mediate the attachment of the virus to host cells and the fusion of viral and cellular membranes. Several membrane factors are involved in virus entry, including C-type lectins and nonmuscle myosin heavy chain IIA. The post-fusion crystal structure of the Gc protein suggests that it is a class II membrane fusion protein, similar to the E/E1 protein of flaviviruses and alphaviruses...
December 16, 2016: Virologica Sinica
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