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Membrane fusion

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https://www.readbyqxmd.com/read/29676464/the-endosomal-recycling-of-fgsnc1-by-fgsnx41-fgsnx4-heterodimer-is-essential-for-polarized-growth-and-pathogenicity-in-fusarium-graminearum
#1
Wenhui Zheng, Yahong Lin, Wenqin Fang, Xu Zhao, Yi Lou, Guanghui Wang, Huawei Zheng, Qifu Liang, Yakubu Saddeeq Abubakar, Stefan Olsson, Jie Zhou, Zonghua Wang
Endosomal sorting machineries regulate the transport of their cargoes among intracellular compartments. However, the molecular nature of such intracellular trafficking processes in pathogenic fungal development and pathogenicity remains unclear. Here, we dissect the roles and molecular mechanisms of two sorting nexin proteins and their cargoes in endosomal recycling in Fusarium graminearum using high-resolution microscopy and high-throughput co-immunoprecipitation strategies. We show that the sorting nexins, FgSnx41 and FgSnx4, interact with each other and assemble into a functionally interdependent heterodimer through their respective BAR domains...
April 20, 2018: New Phytologist
https://www.readbyqxmd.com/read/29675800/secretion-expression-of-human-neutrophil-peptide-1-hnp1-in-pichia-pastoris-and-its-functional-analysis-against-antibiotic-resistant-helicobacter-pylori
#2
Xiaolin Zhang, Anmin Jiang, Banghua Qi, Hao Yu, Youyi Xiong, Guoliang Zhou, Meisong Qin, Jinfeng Dou, Jianfei Wang
Human neutrophil peptide 1 (HNP1) is a small (3.44 kDa) cationic peptide that is a distinct member of the defensin family. HNP1 plays a crucial role in controlling bacterial infections, particularly by antibiotic-resistant bacteria, through membrane perforation patterns. The structural characteristics of HNP1's three intramolecular disulfide bridges cause difficulty in its synthesis via chemical methods. In this study, bioactive recombinant HNP1 was produced using the Pichia pastoris (P. Pichia) expression system...
April 19, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29674345/quantitative-interaction-analysis-permits-molecular-insights-into-functional-nox4-nadph-oxidase-heterodimer-assembly
#3
Sharon O'Neill, Magalie Mathis, Lidija Kovacic, Suisheng Zhang, Juergen Reinhardt, Dimitri Scholz, Ulrich Schopfer, Rochdi Bouhelal, Ulla G Knaus
Protein-protein interactions critically regulate many biological systems, but quantifying functional assembly of multi-pass membrane complexes in their native context is still challenging. Here, we combined modeling-assisted protein modification and information from human disease variants with a minimal-size fusion tag, split-luciferase-based approach to probe assembly of the NADPH oxidase 4 (NOX4)-p22phox enzyme, an integral membrane complex with unresolved structure, which is required for electron transfer and generation of reactive oxygen species (ROS)...
April 19, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29673648/s6-kinase-1-plays-a-key-role-in-mitochondrial-morphology-and-cellular-energy-flow
#4
Quangdon Tran, Jae-Hun Jung, Jisoo Park, Hyunji Lee, Youngeun Hong, Hyeonjeong Cho, Minhee Kim, Sungjin Park, So-Hee Kwon, Seon-Hwan Kim, George Thomas, Kwang Pyo Kim, Myung-Haing Cho, Jongsun Park
Mitochondrial morphology, which is associated with changes in metabolism, cell cycle, cell development and cell death, is tightly regulated by the balance between fusion and fission. In this study, we found that S6 kinase 1 (S6K1) contributes to mitochondrial dynamics, homeostasis and function. Mouse embryo fibroblasts lacking S6K1 (S6K1-KO MEFs) exhibited more fragmented mitochondria and a higher level of Dynamin related protein 1 (Drp1) and active Drp1 (pS616) in both whole cell extracts and mitochondrial fraction...
April 16, 2018: Cellular Signalling
https://www.readbyqxmd.com/read/29671919/a-conserved-rxlr-effector-interacts-with-host-raba-type-gtpases-to-inhibit-vesicle-mediated-secretion-of-antimicrobial-proteins
#5
Iga Tomczynska, Michael Stumpe, Felix Mauch
Plant pathogens of the oomycete genus Phytophthora produce virulence factors, known as RxLR effector proteins that are transferred into host cells to suppress disease resistance. Here, we analyse the function of the highly conserved RxLR24 effector of Phytophthora brassicae. RxLR24 was expressed early in the interaction with Arabidopsis plants and ectopic expression in the host enhanced leaf colonization and zoosporangia formation. Co-immunoprecipitation experiments followed by mass spectrometry identified different members of the RABA GTPase family as putative RxLR24 targets...
April 19, 2018: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/29671763/cd9-and-cd81-interactions-and-their-structural-modelling-in-sperm-prior-to-fertilization
#6
Michaela Frolikova, Pavla Manaskova-Postlerova, Jiri Cerny, Jana Jankovicova, Ondrej Simonik, Alzbeta Pohlova, Petra Secova, Jana Antalikova, Katerina Dvorakova-Hortova
Proteins CD9 and CD81 are members of the tetraspanin superfamily and were detected in mammalian sperm, where they are suspected to form an active tetraspanin web and to participate in sperm⁻egg membrane fusion. The importance of these two proteins during the early stages of fertilization is supported by the complete sterility of CD9/CD81 double null female mice. In this study, the putative mechanism of CD9/CD81 involvement in tetraspanin web formation in sperm and its activity prior to fertilization was addressed...
April 19, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29671361/liposomal-quercetin-potentiates-maxi-k-channel-openings-in-smooth-muscles-and-restores-its-activity-after-oxidative-stress
#7
Mariia I Melnyk, Dariia O Dryn, Lina T Al Kury, Alexander V Zholos, Anatoly I Soloviev
The effects of quercetin-loaded liposomes (PCL-Q) and their constituents, that is, free quercetin (Q) and 'empty' phosphatidylcholine vesicles (PCL), on maxi-K channel activity were studied in single mouse ileal myocytes before and after H2 O2 -induced oxidative stress. Macroscopic Maxi-K channel currents were recorded using whole-cell patch clamp techniques, while single BKCa channel currents were recorded in the cell-attached configuration. Bath application of PCL-Q (100 μg/ml of lipid and 3 μg/ml of quercetin) increased single Maxi-K channel activity more than threefold, from 0...
April 19, 2018: Journal of Liposome Research
https://www.readbyqxmd.com/read/29669920/reduced-vacuolar-%C3%AE-1-3-glucan-synthesis-affects-carbohydrate-metabolism-as-well-as-plastid-homeostasis-and-structure-in-phaeodactylum-tricornutum
#8
Weichao Huang, Ilka Haferkamp, Bernard Lepetit, Mariia Molchanova, Shengwei Hou, Wolfgang Jeblick, Carolina Río Bártulos, Peter G Kroth
The β-1,3-glucan chrysolaminarin is the main storage polysaccharide of diatoms. In contrast to plants and green algae, diatoms and most other algal groups do not accumulate storage polysaccharides in their plastids. The diatom Phaeodactylum tricornutum possesses only a single gene encoding a putative β-1,3-glucan synthase ( Pt BGS). Here, we characterize this enzyme by expressing GFP fusion proteins in P. tricornutum and by creating and investigating corresponding gene silencing mutants. We demonstrate that Pt BGS is a vacuolar protein located in the tonoplast...
April 18, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29669880/transmembrane-domains-of-highly-pathogenic-viral-fusion-proteins-exhibit-trimeric-association-in-vitro
#9
Stacy R Webb, Stacy E Smith, Michael G Fried, Rebecca Ellis Dutch
Enveloped viruses require viral fusion proteins to promote fusion of the viral envelope with a target cell membrane. To drive fusion, these proteins undergo large conformational changes that must occur at the right place and at the right time. Understanding the elements which control the stability of the prefusion state and the initiation of conformational changes is key to understanding the function of these important proteins. The construction of mutations in the fusion protein transmembrane domains (TMDs) or the replacement of these domains with lipid anchors has implicated the TMD in the fusion process...
April 25, 2018: MSphere
https://www.readbyqxmd.com/read/29669253/an-antibody-targeting-the-fusion-machinery-neutralizes-dual-tropic-infection-and-defines-a-site-of-vulnerability-on-epstein-barr-virus
#10
Joost Snijder, Michael S Ortego, Connor Weidle, Andrew B Stuart, Matthew D Gray, M Juliana McElrath, Marie Pancera, David Veesler, Andrew T McGuire
Epstein-Barr virus (EBV) is a causative agent of infectious mononucleosis and is associated with 200,000 new cases of cancer and 140,000 deaths annually. Subunit vaccines against this pathogen have focused on the gp350 glycoprotein and remain unsuccessful. We isolated human antibodies recognizing the EBV fusion machinery (gH/gL and gB) from rare memory B cells. One anti-gH/gL antibody, AMMO1, potently neutralized infection of B cells and epithelial cells, the two major cell types targeted by EBV. We determined a cryo-electron microscopy reconstruction of the gH/gL-gp42-AMMO1 complex and demonstrated that AMMO1 bound to a discontinuous epitope formed by both gH and gL at the Domain-I/Domain-II interface...
April 17, 2018: Immunity
https://www.readbyqxmd.com/read/29668757/actin-polymerization-in-the-endosomal-pathway-but-not-on-the-coxiella-containing-vacuole-is-essential-for-pathogen-growth
#11
Heather E Miller, Charles L Larson, Robert A Heinzen
Coxiella burnetii is an intracellular bacterium that replicates within an expansive phagolysosome-like vacuole. Fusion between the Coxiella-containing vacuole (CCV) and late endosomes/multivesicular bodies requires Rab7, the HOPS tethering complex, and SNARE proteins, with actin also speculated to play a role. Here, we investigated the importance of actin in CCV fusion. Filamentous actin patches formed around the CCV membrane that were preferred sites of vesicular fusion. Accordingly, the mediators of endolysosomal fusion Rab7, VAMP7, and syntaxin 8 were concentrated in CCV actin patches...
April 18, 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/29667836/molecular-dynamics-simulations-are-redefining-our-view-of-peptides-interacting-with-biological-membranes
#12
Jakob P Ulmschneider, Martin B Ulmschneider
Ever since the first molecular mechanics computer simulations of biological molecules became possible, there has been the dream to study all complex biological phenomena in silico, simply bypassing the enormous experimental challenges and their associated costs. For this, two inherent requirements need to be met: First, the time scales achievable in simulations must reach up to the millisecond range and even longer. Second, the computational model must accurately reproduce what is measured experimentally. Despite some recent successes, the general consensus in the field to date has been that neither of these conditions have yet been met and that the dream will be realized, if at all, only in the distant future...
April 18, 2018: Accounts of Chemical Research
https://www.readbyqxmd.com/read/29667660/a-new-functional-membrane-protein-microarray-based-on-tethered-phospholipid-bilayers
#13
Meriem Chadli, Ofelia Maniti, Christophe Marquette, Bruno Tillier, Sandra Cortès, Agnès Girard-Egrot
A new prototype of a membrane protein biochip is presented in this article. This biochip was created by the combination of novel technologies of peptide-tethered bilayer lipid membrane (pep-tBLM) formation and solid support micropatterning. Pep-tBLMs integrating a membrane protein were obtained in the form of microarrays on a gold chip. The formation of the microspots was visualized in real-time by surface plasmon resonance imaging (SPRi) and the functionality of a GPCR (CXCR4), reinserted locally into microwells, was assessed by ligand binding studies...
April 18, 2018: Analyst
https://www.readbyqxmd.com/read/29667004/the-development-of-hiv-vaccines-targeting-gp41-membrane-proximal-external-region-mper-challenges-and-prospects
#14
REVIEW
Huan Liu, Xiaojie Su, Lulu Si, Lu Lu, Shibo Jiang
A human immunodeficiency virus type-1 (HIV-1) vaccine which is able to effectively prevent infection would be the most powerful method of extinguishing pandemic of the acquired immunodeficiency syndrome (AIDS). Yet, achieving such vaccine remains great challenges. The membrane-proximal external region (MPER) is a highly conserved region of the envelope glycoprotein (Env) gp41 subunit near the viral envelope surface, and it plays a key role in membrane fusion. It is also the target of some reported broadly neutralizing antibodies (bNAbs)...
April 17, 2018: Protein & Cell
https://www.readbyqxmd.com/read/29666192/arrest-of-trans-snare-zippering-uncovers-loosely-and-tightly-docked-intermediates-in-membrane-fusion
#15
Halenur Yavuz, Iman Kattan, Javier Matias Hernandez, Oliver Hofnagel, Agata Witkowska, Stefan Raunser, Peter Jomo Walla, Reinhard Jahn
Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediate intracellular membrane fusion in the secretory pathway. They contain conserved regions, termed SNARE motifs, that assemble between opposing membranes directionally from their N-termini to their membrane-proximal C-termini in a highly exergonic reaction. However, how this energy is utilized to overcome the energy barriers along the fusion pathway is still under debate. Here we have used mutants of the SNARE synaptobrevin to arrest trans-SNARE zippering at defined stages...
April 17, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29664647/amphiphobic-septa-enhance-the-mechanical-stability-of-free-standing-bilayer-lipid-membranes
#16
Daichi Yamaura, Daisuke Tadaki, Shun Araki, Miyu Yoshida, Kohei Arata, Takeshi Ohori, Ken-Ichi Ishibashi, Miki Kato, Teng Ma, Ryusuke Miyata, Hideaki Yamamoto, Ryugo Tero, Masao Sakuraba, Toshio Ogino, Michio Niwano, Ayumi Hirano-Iwata
Artificial bilayer lipid membranes (BLMs) provide well-defined systems for investigating the fundamental properties of membrane proteins, including ion channels, and in screening the effect of drugs that act on them. However, the application of this technique is limited due to the low stability and low reconstitution efficiency of the process. We previously reported on improving the stability of BLM based on the fabrication of microapertures having a tapered edge in SiO2/Si3N4 septa and efficient ion channel incorporation based on vesicle fusion accelerated by a centrifugal force...
April 17, 2018: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/29663589/tying-trafficking-to-fusion-and-fission-at-the-mighty-mitochondria
#17
REVIEW
Trey Farmer, Naava Naslavsky, Steve Caplan
The mitochondrion is a unique organelle that serves as the main site of adenosine triphosphate (ATP) generation needed for energy in the cell. However, mitochondria also play essential roles in cell death through apoptosis and necrosis, as well as a variety of crucial functions related to stress regulation, autophagy, lipid synthesis, and calcium storage. There is a growing appreciation that mitochondrial function is regulated by the dynamics of its membrane fusion and fission; longer, fused mitochondria are optimal for ATP generation, whereas fission of mitochondria facilitates mitophagy and cell division...
April 16, 2018: Traffic
https://www.readbyqxmd.com/read/29663420/phase-1-first-in-human-single-and-multiple-ascending-dose-and-food-effect-studies-to-assess-the-safety-tolerability-and-pharmacokinetics-of-presatovir-for-the-treatment-of-respiratory-syncytial-virus-infection
#18
Polina German, Yan Xin, Jason W Chien, Winnie Weng, Richard Mackman, Sandra A Lewis, Amy Meng, John Ling, Anita Mathias
Respiratory syncytial virus (RSV)-associated respiratory tract infection is a leading cause of hospitalizations in infants for which no effective treatment exists. RSV infection is also an important cause of respiratory disease in adults and immunocompromised patients. Presatovir (GS-5806) is an orally bioavailable antiviral agent that inhibits fusion of RSV with host cell membranes. Here, results from 2 phase 1 studies that evaluated safety, tolerability, and pharmacokinetics of presatovir in healthy adults following administration of single and multiple (7 days) once- or twice-daily ascending doses (first-in-human study) and in the presence or absence of food (food effect study) are described...
April 17, 2018: Journal of Clinical Pharmacology
https://www.readbyqxmd.com/read/29662028/nitrate-reductases-are-relocalized-to-the-nucleus-by-atsiz1-and-their-levels-are-negatively-regulated-by-cop1-and-ammonium
#19
Joo Yong Kim, Bong Soo Park, Sang Woo Park, Han Yong Lee, Jong Tae Song, Hak Soo Seo
Nitrate reductases (NRs) catalyze the first step in the reduction of nitrate to ammonium. NR activity is regulated by sumoylation through the E3 ligase activity of AtSIZ1. However, it is not clear how NRs interact with AtSIZ1 in the cell, or how nitrogen sources affect NR levels and their cellular localization. Here, we show that the subcellular localization of NRs is modulated by the E3 SUMO (Small ubiquitin-related modifier) ligase AtSIZ1 and that NR protein levels are regulated by nitrogen sources. Transient expression analysis of GFP fusion proteins in onion epidermal cells showed that the NRs NIA1 and NIA2 localize to the cytoplasmic membrane, and that AtSIZ1 localizes to the nucleoplasm, including nuclear bodies, when expressed separately, whereas NRs and AtSIZ1 localize to the nucleus when co-expressed...
April 15, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29661855/the-heptad-repeat-domain-1-of-mitofusin-has-membrane-destabilization-function-in-mitochondrial-fusion
#20
Frédéric Daste, Cécile Sauvanet, Andrej Bavdek, James Baye, Fabienne Pierre, Rémi Le Borgne, Claudine David, Manuel Rojo, Patrick Fuchs, David Tareste
Mitochondria are double-membrane-bound organelles that constantly change shape through membrane fusion and fission. Outer mitochondrial membrane fusion is controlled by Mitofusin, whose molecular architecture consists of an N-terminal GTPase domain, a first heptad repeat domain (HR1), two transmembrane domains, and a second heptad repeat domain (HR2). The mode of action of Mitofusin and the specific roles played by each of these functional domains in mitochondrial fusion are not fully understood. Here, using a combination of in situ and in vitro fusion assays, we show that HR1 induces membrane fusion and possesses a conserved amphipathic helix that folds upon interaction with the lipid bilayer surface...
April 16, 2018: EMBO Reports
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