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Crystallization of membrane protein

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https://www.readbyqxmd.com/read/27909292/crystal-structures-of-the-tric-trimeric-intracellular-cation-channel-orthologues
#1
Go Kasuya, Masahiro Hiraizumi, Andrés D Maturana, Kaoru Kumazaki, Yuichiro Fujiwara, Keihong Liu, Yoshiko Nakada-Nakura, So Iwata, Keisuke Tsukada, Tomotaka Komori, Sotaro Uemura, Yuhei Goto, Takanori Nakane, Mizuki Takemoto, Hideaki E Kato, Keitaro Yamashita, Miki Wada, Koichi Ito, Ryuichiro Ishitani, Motoyuki Hattori, Osamu Nureki
Ca(2+) release from the sarcoplasmic reticulum (SR) and endoplasmic reticulum (ER) is crucial for muscle contraction, cell growth, apoptosis, learning and memory. The trimeric intracellular cation (TRIC) channels were recently identified as cation channels balancing the SR and ER membrane potentials, and are implicated in Ca(2+) signaling and homeostasis. Here we present the crystal structures of prokaryotic TRIC channels in the closed state and structure-based functional analyses of prokaryotic and eukaryotic TRIC channels...
December 2016: Cell Research
https://www.readbyqxmd.com/read/27907120/structural-and-functional-elucidation-of-yeast-lanosterol-14%C3%AE-demethylase-in-complex-with-agrochemical-antifungals
#2
Joel D A Tyndall, Manya Sabherwal, Alia A Sagatova, Mikhail V Keniya, Jacopo Negroni, Rajni K Wilson, Matthew A Woods, Klaus Tietjen, Brian C Monk
Azole antifungals, known as demethylase inhibitors (DMIs), target sterol 14α-demethylase (CYP51) in the ergosterol biosynthetic pathway of fungal pathogens of both plants and humans. DMIs remain the treatment of choice in crop protection against a wide range of fungal phytopathogens that have the potential to reduce crop yields and threaten food security. We used a yeast membrane protein expression system to overexpress recombinant hexahistidine-tagged S. cerevisiae lanosterol 14α-demethylase and the Y140F or Y140H mutants of this enzyme as surrogates in order characterize interactions with DMIs...
2016: PloS One
https://www.readbyqxmd.com/read/27906127/malaria-parasite-celtos-targets-the-inner-leaflet-of-cell-membranes-for-pore-dependent-disruption
#3
John R Jimah, Nichole D Salinas, Monica Sala-Rabanal, Nathaniel G Jones, L David Sibley, Colin G Nichols, Paul H Schlesinger, Niraj H Tolia
Apicomplexan parasites contain a conserved protein CelTOS that, in malaria parasites, is essential for traversal of cells within the mammalian host and arthropod vector. However, the molecular role of CelTOS is unknown because it lacks sequence similarity to proteins of known function. Here, we determined the crystal structure of CelTOS and discovered CelTOS resembles proteins that bind to and disrupt membranes. In contrast to known membrane disruptors, CelTOS has a distinct architecture, specifically binds phosphatidic acid commonly present within the inner leaflet of plasma membranes, and potently disrupts liposomes composed of phosphatidic acid by forming pores...
December 1, 2016: ELife
https://www.readbyqxmd.com/read/27899666/structures-of-human-srp72-complexes-provide-insights-into-srp-rna-remodeling-and-ribosome-interaction
#4
Matthias M M Becker, Karine Lapouge, Bernd Segnitz, Klemens Wild, Irmgard Sinning
Co-translational protein targeting and membrane protein insertion is a fundamental process and depends on the signal recognition particle (SRP). In mammals, SRP is composed of the SRP RNA crucial for SRP assembly and function and six proteins. The two largest proteins SRP68 and SRP72 form a heterodimer and bind to a regulatory site of the SRP RNA. Despite their essential roles in the SRP pathway, structural information has been available only for the SRP68 RNA-binding domain (RBD). Here we present the crystal structures of the SRP68 protein-binding domain (PBD) in complex with SRP72-PBD and of the SRP72-RBD bound to the SRP S domain (SRP RNA, SRP19 and SRP68) detailing all interactions of SRP72 within SRP...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27883993/stimulation-of-the-nonneuronal-cholinergic-system-by-highly-diluted-acetylcholine-in-keratinocytes
#5
Francesca Uberti, Claudio Bardelli, Vera Morsanuto, Sabrina Ghirlanda, Andrea Cochis, Claudio Molinari
The physiological effects of acetylcholine on keratinocytes depend on the presence of nicotinic and muscarinic receptors. The role of nonneuronal acetylcholine in keratinocytes could have important clinical implications for patients with various skin disorders such as nonhealing wounds. In order to evaluate the efficacy of highly diluted acetylcholine solutions obtained by sequential kinetic activation, we aimed to investigate the effects of these solutions on normal human keratinocytes. Two different concentrations (10 fg/mL and 1 pg/mL) and formulations (kinetically activated and nonkinetically activated) of acetylcholine were used to verify keratinocyte viability, proliferation, and migration and the intracellular pathways involved using MTT, crystal violet, wound healing, and Western blot compared to 147 ng/mL acetylcholine...
November 25, 2016: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/27883039/structural-and-functional-probing-of-porz-an-essential-bacterial-surface-component-of-the-type-ix-secretion-system-of-human-oral-microbiomic-porphyromonas-gingivalis
#6
Anna M Lasica, Theodoros Goulas, Danuta Mizgalska, Xiaoyan Zhou, Iñaki de Diego, Mirosław Ksiazek, Mariusz Madej, Yonghua Guo, Tibisay Guevara, Magdalena Nowak, Barbara Potempa, Apoorv Goel, Maryta Sztukowska, Apurva T Prabhakar, Monika Bzowska, Magdalena Widziolek, Ida B Thøgersen, Jan J Enghild, Mary Simonian, Arkadiusz W Kulczyk, Ky-Anh Nguyen, Jan Potempa, F Xavier Gomis-Rüth
Porphyromonas gingivalis is a member of the human oral microbiome abundant in dysbiosis and implicated in the pathogenesis of periodontal (gum) disease. It employs a newly described type-IX secretion system (T9SS) for secretion of virulence factors. Cargo proteins destined for secretion through T9SS carry a recognition signal in the conserved C-terminal domain (CTD), which is removed by sortase PorU during translocation. Here, we identified a novel component of T9SS, PorZ, which is essential for surface exposure of PorU and posttranslational modification of T9SS cargo proteins...
November 24, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27881302/crystal-structure-of-a-full-length-human-tetraspanin-reveals-a-cholesterol-binding-pocket
#7
Brandon Zimmerman, Brendan Kelly, Brian J McMillan, Tom C M Seegar, Ron O Dror, Andrew C Kruse, Stephen C Blacklow
Tetraspanins comprise a diverse family of four-pass transmembrane proteins that play critical roles in the immune, reproductive, genitourinary, and auditory systems. Despite their pervasive roles in human physiology, little is known about the structure of tetraspanins or the molecular mechanisms underlying their various functions. Here, we report the crystal structure of human CD81, a full-length tetraspanin. The transmembrane segments of CD81 pack as two largely separated pairs of helices, capped by the large extracellular loop (EC2) at the outer membrane leaflet...
November 3, 2016: Cell
https://www.readbyqxmd.com/read/27879668/structure-functional-basis-of-ion-transport-in-sodium-calcium-exchanger-ncx-proteins
#8
REVIEW
Moshe Giladi, Reut Shor, Michal Lisnyansky, Daniel Khananshvili
The membrane-bound sodium-calcium exchanger (NCX) proteins shape Ca(2+) homeostasis in many cell types, thus participating in a wide range of physiological and pathological processes. Determination of the crystal structure of an archaeal NCX (NCX_Mj) paved the way for a thorough and systematic investigation of ion transport mechanisms in NCX proteins. Here, we review the data gathered from the X-ray crystallography, molecular dynamics simulations, hydrogen-deuterium exchange mass-spectrometry (HDX-MS), and ion-flux analyses of mutants...
November 22, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27872308/atomistic-insight-into-lipid-translocation-by-a-tmem16-scramblase
#9
Neville P Bethel, Michael Grabe
The transmembrane protein 16 (TMEM16) family of membrane proteins includes both lipid scramblases and ion channels involved in olfaction, nociception, and blood coagulation. The crystal structure of the fungal Nectria haematococca TMEM16 (nhTMEM16) scramblase suggested a putative mechanism of lipid transport, whereby polar and charged lipid headgroups move through the low-dielectric environment of the membrane by traversing a hydrophilic groove on the membrane-spanning surface of the protein. Here, we use computational methods to explore the membrane-protein interactions involved in lipid scrambling...
November 21, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27872187/galactomannan-catabolism-conferred-by-a-polysaccharide-utilisation-locus-of-bacteroides-ovatus-enzyme-synergy-and-crystal-structure-of-a-%C3%AE-mannanase
#10
Viktoria Bågenholm, Sumitha K Reddy, Hanene Bouraoui, Johan Morrill, Evelina Kulcinskaja, Constance M Bahr, Oskar Aurelius, Theresa Rogers, Yao Xiao, Derek T Logan, Eric C Martens, Nicole M Koropatkin, Henrik Stålbrand
A recently identified polysaccharide utilisation locus (PUL) from Bacteroides ovatus ATCC8483 is transcriptionally upregulated during growth on galacto- and glucomannans. It encodes two putative glycoside hydrolase family 26 (GH26) β-mannanases, BoMan26A and BoMan26B, and a GH36 α-galactosidase, BoGal36A. The PUL also includes two glycan binding proteins, confirmed by β-mannan affinity electrophoresis. When this PUL was deleted, B. ovatus was no longer able to grow on locust bean galactomannan. BoMan26A primarily formed mannobiose from mannan polysaccharides...
November 21, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27870886/comparative-genomics-revealed-multiple-helicobacter-pylori-genes-associated-with-biofilm-formation-in-vitro
#11
Eric Hong Jian Wong, Chow Goon Ng, Eng Guan Chua, Alfred Chin Yen Tay, Fanny Peters, Barry J Marshall, Bow Ho, Khean Lee Goh, Jamuna Vadivelu, Mun Fai Loke
BACKGROUND: Biofilm formation by Helicobacter pylori may be one of the factors influencing eradication outcome. However, genetic differences between good and poor biofilm forming strains have not been studied. MATERIALS AND METHODS: Biofilm yield of 32 Helicobacter pylori strains (standard strain and 31 clinical strains) were determined by crystal-violet assay and grouped into poor, moderate and good biofilm forming groups. Whole genome sequencing of these 32 clinical strains was performed on the Illumina MiSeq platform...
2016: PloS One
https://www.readbyqxmd.com/read/27859784/membrane-skeletal-association-and-post-translational-allosteric-regulation-of-toxoplasma-gondii-gapdh1
#12
Rashmi Dubey, Bart L Staker, Ian T Foe, Matthew Bogyo, Peter J Myler, Huân M Ngô, Marc-Jan Gubbels
When Toxoplasma gondii egresses from the host cell, glyceraldehyde-3-phosphate dehydrogenase 1 (GAPDH1), which is primary a glycolysis enzyme but actually a quintessential multifunctional protein, translocates to the unique cortical membrane skeleton. Here we report the 2.25Å resolution crystal structure of the GAPDH1 holoenzyme in a quaternary complex providing the basis for the molecular dissection of GAPDH1 structure-function relationships Knockdown of GAPDH1 expression and catalytic site disruption validate the essentiality of GAPDH1 in intracellular replication but we confirmed that glycolysis is not strictly essential...
November 17, 2016: Molecular Microbiology
https://www.readbyqxmd.com/read/27851727/high-resolution-crystal-structure-of-the-human-cb1-cannabinoid-receptor
#13
Zhenhua Shao, Jie Yin, Karen Chapman, Magdalena Grzemska, Lindsay Clark, Junmei Wang, Daniel M Rosenbaum
The human cannabinoid G-protein-coupled receptors (GPCRs) CB1 and CB2 mediate the functional responses to the endocannabinoids anandamide and 2-arachidonyl glycerol (2-AG), as well as the widely consumed plant (phyto)cannabinoid Δ(9)-tetrahydrocannabinol (THC)(1). The cannabinoid receptors have been the targets of intensive drug discovery efforts owing to the therapeutic potential of modulators for controlling pain(2), epilepsy(3), obesity(4), and other maladies. Although much progress has recently been made in understanding the biophysical properties of GPCRs, investigations of the molecular mechanisms of the cannabinoids and their receptors have lacked high-resolution structural data...
November 16, 2016: Nature
https://www.readbyqxmd.com/read/27832483/7tm-domain-structure-of-adhesion-gpcrs
#14
Chris de Graaf, Saskia Nijmeijer, Steffen Wolf, Oliver P Ernst
Schematic presentation of the overall adhesion G Protein-Coupled Receptor (aGPCR) structure and functional domains, covering an extracellular N-terminal fragment (NTF), a membrane-spanning C-terminal fragment (CTF) and a GPCR proteolysis site (GPS). (Left side) aGPCR model constructed based on the seven-transmembrane (7TM) structure (blue) of secretin family glucagon receptor (GCGR) (PDB, 4L6R) [11] and the GPCR autoproteolysis inducing (GAIN) domain (magenta) structure of latrophilin 1 (PDB, 4DLQ) [9]...
2016: Handbook of Experimental Pharmacology
https://www.readbyqxmd.com/read/27832149/the-structure-of-treponema-pallidum-tp0624-reveals-a-modular-assembly-of-divergently-functionalized-and-previously-uncharacterized-domains
#15
Michelle L Parker, Simon Houston, Charmaine Wetherell, Caroline E Cameron, Martin J Boulanger
Treponema pallidum subspecies pallidum is the causative agent of syphilis, a chronic, multistage, systemic infection that remains a major global health concern. The molecular mechanisms underlying T. pallidum pathogenesis are incompletely understood, partially due to the phylogenetic divergence of T. pallidum. One aspect of T. pallidum that differentiates it from conventional Gram-negative bacteria, and is believed to play an important role in pathogenesis, is its unusual cell envelope ultrastructure; in particular, the T...
2016: PloS One
https://www.readbyqxmd.com/read/27830695/crystal-structure-of-bacterial-haem-importer-complex-in-the-inward-facing-conformation
#16
Youichi Naoe, Nozomi Nakamura, Akihiro Doi, Mia Sawabe, Hiro Nakamura, Yoshitsugu Shiro, Hiroshi Sugimoto
Pathogenic bacteria remove iron from the haem of host tissues and use it as a catalytic center of many enzymes. Haem uptake by pathogenic bacteria is facilitated by the membrane-integrated haem importer, which belongs to the type II ATP-binding cassette (ABC) transporter. Here we present crystal structures of Burkholderia cenocepacia haem importer BhuUV complexed with the periplasmic haem-binding protein BhuT and in the absence of BhuT. The transmembrane helices of these structures show an inward-facing conformation, in which the cytoplasmic gate of the haem translocation pathway is completely open...
November 10, 2016: Nature Communications
https://www.readbyqxmd.com/read/27827364/structural-and-mechanistic-insights-into-regulation-of-the-retromer-coat-by-tbc1d5
#17
Da Jia, Jin-San Zhang, Fang Li, Jing Wang, Zhihui Deng, Mark A White, Douglas G Osborne, Christine Phillips-Krawczak, Timothy S Gomez, Haiying Li, Amika Singla, Ezra Burstein, Daniel D Billadeau, Michael K Rosen
Retromer is a membrane coat complex that is recruited to endosomes by the small GTPase Rab7 and sorting nexin 3. The timing of this interaction and consequent endosomal dynamics are thought to be regulated by the guanine nucleotide cycle of Rab7. Here we demonstrate that TBC1d5, a GTPase-activating protein (GAP) for Rab7, is a high-affinity ligand of the retromer cargo selective complex VPS26/VPS29/VPS35. The crystal structure of the TBC1d5 GAP domain bound to VPS29 and complementary biochemical and cellular data show that a loop from TBC1d5 binds to a conserved hydrophobic pocket on VPS29 opposite the VPS29-VPS35 interface...
November 9, 2016: Nature Communications
https://www.readbyqxmd.com/read/27825102/microvillar-injury-in-renal-tubular-epithelial-cells-induced-by-calcium-oxalate-crystal-and-the-protective-role-of-epigallocatechin-3-gallate
#18
Kedsarin Fong-Ngern, Arada Vinaiphat, Visith Thongboonkerd
Pathogenic mechanisms of kidney stone disease remained unclear. This study investigated its initial cellular/molecular mechanisms when calcium oxalate monohydrate (COM) crystal adhered to renal tubular cells. Transmission electron microscopy revealed decreased length and density of microvilli, whereas Western blot analysis showed that whole-cell ezrin (a microvilli-stabilizing protein), not β-actin, was decreased in COM-treated cells. Immunofluorescence staining, followed by laser-scanning confocal microscopy and subcellular fractionations, revealed decreases in both ezrin and F-/β-actin at apical membrane...
October 7, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/27822186/structure-and-protein-protein-interactions-of-human-udp-glucuronosyltransferases
#19
REVIEW
Ryoichi Fujiwara, Tsuyoshi Yokoi, Miki Nakajima
Mammalian UDP-glucuronosyltransferases (UGTs) catalyze the transfer of glucuronic acid from UDP-glucuronic acid to various xenobiotics and endobiotics. Since UGTs comprise rate-limiting enzymes for metabolism of various compounds, co-administration of UGT-inhibiting drugs and genetic deficiency of UGT genes can cause an increased blood concentration of these compounds. During the last few decades, extensive efforts have been made to advance the understanding of gene structure, function, substrate specificity, and inhibition/induction properties of UGTs...
2016: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/27821762/x-ray-and-cryo-em-structures-of-monomeric-and-filamentous-actin-like-protein-mamk-reveal-changes-associated-with-polymerization
#20
Jan Löwe, Shaoda He, Sjors H W Scheres, Christos G Savva
Magnetotactic bacteria produce iron-rich magnetic nanoparticles that are enclosed by membrane invaginations to form magnetosomes so they are able to sense and act upon Earth's magnetic field. In Magnetospirillum and other magnetotactic bacteria, to combine their magnetic moments, magnetosomes align along filaments formed by a bacterial actin homolog, MamK. Here, we present the crystal structure of a nonpolymerizing mutant of MamK from Magnetospirillum magneticum AMB-1 at 1.8-Å resolution, revealing its close similarity to actin and MreB...
November 7, 2016: Proceedings of the National Academy of Sciences of the United States of America
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