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https://www.readbyqxmd.com/read/28545444/site-specific-his-asp-phosphoproteomic-analysis-of-prokaryotes-reveals-putative-targets-for-drug-resistance
#1
Shu-Jung Lai, I-Fan Tu, Wan-Ling Wu, Jhih-Tian Yang, Louis Y P Luk, Mei-Chin Lai, Yu-Hsuan Tsai, Shih-Hsiung Wu
BACKGROUND: Phosphorylation of amino acid residues on proteins is an important and common post-translational modification in both eukaryotes and prokaryotes. Most research work has been focused on phosphorylation of serine, threonine or tyrosine residues, whereas phosphorylation of other amino acids are significantly less clear due to the controversy on their stability under standard bioanalytical conditions. RESULTS: Here we applied a shotgun strategy to analyze the histidine and aspartate phosphorylations in different microbes...
May 25, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28544379/a-widened-substrate-selectivity-filter-of-eukaryotic-formate-nitrite-transporters-enables-high-level-lactate-conductance
#2
Marie Wiechert, Holger Erler, André Golldack, Eric Beitz
Bacterial formate-nitrite transporters (FNT) regulate the metabolic flow of small weak mono-acids derived from anaerobic mixed-acid fermentation, such as formate, and further transport nitrite, and hydrosulfide. The eukaryotic Plasmodium falciparum FNT is vital for the malaria parasite by its ability to release the larger l-lactate substrate as the metabolic end product of anaerobic glycolysis in symport with protons preventing cytosolic acidification. However, the molecular basis for substrate discrimination by FNTs has remained unclear...
May 21, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28544098/crystal-structure-of-the-sensor-domain-of-baes-from-serratia-marcescens-fs14
#3
Ya Zhang, Shenshen Qiu, Shanshan Jia, Dongqing Xu, Tingting Ran, Weiwu Wang
The sensor histidine kinases of two-component signal-transduction systems (TCSs) are essential for bacteria to adapt to variable environmental conditions. The two-component regulatory system BaeS/R increases multidrug and metal resistance in Salmonella and Escherichia coli. In this study, we report the X-ray structure of the periplasmic sensor domain of BaeS from Serratia marcescens FS14. The BaeS sensor domain (34-160) adopts a mixed α/β-fold containing a central four-stranded antiparallel β-sheet flanked by a long N-terminal α-helix and additional loops and a short C-terminal α-helix on each side...
May 24, 2017: Proteins
https://www.readbyqxmd.com/read/28542513/structural-features-of-phox-one-of-the-phosphate-binding-proteins-from-pho-regulon-of-xanthomonas-citri
#4
Vanessa R Pegos, Rodrigo M L Santos, Francisco J Medrano, Andrea Balan
In Escherichia coli, the ATP-Binding Cassette transporter for phosphate is encoded by the pstSCAB operon. PstS is the periplasmic component responsible for affinity and specificity of the system and has also been related to a regulatory role and chemotaxis during depletion of phosphate. Xanthomonas citri has two phosphate-binding proteins: PstS and PhoX, which are differentially expressed under phosphate limitation. In this work, we focused on PhoX characterization and comparison with PstS. The PhoX three-dimensional structure was solved in a closed conformation with a phosphate engulfed in the binding site pocket between two domains...
2017: PloS One
https://www.readbyqxmd.com/read/28539918/the-sapa-protein-is-involved-in-resistance-to-antimicrobial-peptide-pr-39-and-virulence-of-actinobacillus-pleuropneumoniae
#5
Fang Xie, Yalei Wang, Gang Li, Shuanghong Liu, Ning Cui, Siguo Liu, Paul R Langford, Chunlai Wang
Antimicrobial peptides are essential to the innate immune defense of the mammal against bacterial infection. However, pathogenic bacteria have evolved multiple strategies to resist and evade antimicrobial peptides, which is vital to bacterial survival and colonization in hosts. PR-39 is a linear porcine antimicrobial peptide containing 39 amino acid residues with a high proline content. Resistance to antimicrobial peptide PR-39 has been observed in Actinobacillus pleuropneumoniae. However, little is known about the factors required for this resistance...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28539041/outer-membrane-vesicles-derived-from-salmonella-enteritidis-protect-against-the-virulent-wild-type-strain-infection-in-a-mouse-model
#6
Qiong Liu, Jie Yi, Kang Liang, Xiangmin Zhang, Qing Liu
Food-borne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)...
May 25, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28538146/ligand-mediated-folding-of-the-ompa-periplasmic-domain-from-acinetobacter-baumannii
#7
Ameeq Ul Mushtaq, Jeong Soon Park, Sung-Hun Bae, Hye-Yeon Kim, Kwon Joo Yeo, Eunha Hwang, Ki Yong Lee, Jun-Goo Jee, Hae-Kap Cheong, Young Ho Jeon
The periplasmic domain of OmpA from Acinetobacter baumannii (AbOmpA-PD) binds to diaminopimelate and anchors the outer membrane to the peptidoglycan layer in the cell wall. Although the crystal structure of AbOmpA-PD with its ligands has been reported, the mechanism of ligand-mediated folding of AbOmpA remains elusive. Here, we report that in vitro refolded apo-AbOmpA-PD in the absence of ligand exists as a mixture of two partially folded forms in solution: mostly unfolded (apo-state I) and hololike (apo-state II) states...
May 23, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28523273/novel-treponema-pallidum-recombinant-antigens-for-syphilis-diagnostics-current-status-and-future-prospects
#8
REVIEW
Aleksey Kubanov, Anastassia Runina, Dmitry Deryabin
The recombinant protein technology considerably promoted the development of rapid and accurate treponema-specific laboratory diagnostics of syphilis infection. For the last ten years, the immunodominant recombinant inner membrane lipoproteins are proved to be sensitive and specific antigens for syphilis screening. However, the development of an enlarged T. pallidum antigen panel for diagnostics of early and late syphilis and differentiation of syphilis stages or cured syphilis remains as actual goal of multidisciplinary expertise...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28522691/mechanism-of-transmembrane-signaling-by-sensor-histidine-kinases
#9
Ivan Gushchin, Igor Melnikov, Vitaly Polovinkin, Andrii Ishchenko, Anastasia Yuzhakova, Pavel Buslaev, Gleb Bourenkov, Sergei Grudinin, Ekaterina Round, Taras Balandin, Valentin Borshchevskiy, Dieter Willbold, Gordon Leonard, Georg Büldt, Alexander Popov, Valentin Gordeliy
One of the major and essential classes of transmembrane (TM) receptors, present in all domains of life, is sensor histidine kinases (HKs), parts of two-component signaling systems (TCS). The structural mechanisms of transmembrane signaling by these sensors are poorly understood. We present here crystal structures of the periplasmic sensor domain, the TM domain and the cytoplasmic HAMP domain of the Escherichia coli nitrate/nitrite sensor HK NarQ in the ligand-bound and mutated ligand-free states. The structures reveal that the ligand binding induces significant rearrangements and piston-like shifts of TM helices...
May 18, 2017: Science
https://www.readbyqxmd.com/read/28515319/the-h-region-of-twin-arginine-signal-peptides-supports-productive-binding-of-bacterial-tat-precursor-proteins-to-the-tatbc-receptor-complex
#10
Agnes Ulfig, Julia Fröbel, Frank Lausberg, Anne-Sophie Blümmel, Anna Katharina Heide, Matthias Müller, Roland Freudl
The twin arginine translocation (Tat) pathway transports folded proteins across bacterial membranes. Tat precursor proteins possess a conserved twin-arginine (RR) motif in their signal peptides that is involved in their binding to the Tat translocase, but some facets of this interaction remain unclear. Here, we investigated the role of the hydrophobic (h-) region of the Escherichia coli TMAO reductase (TorA) signal peptide in TatBC receptor binding in vivo and in vitro. We show that besides the RR motif, a minimal functional h-region in the signal peptide is required for Tat-dependent export in E...
May 17, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28512454/new-insights-into-the-regulation-of-cell-surface-signaling-activity-acquired-from-a-mutagenesis-screen-of-the-pseudomonas-putida-iuty-sigma-anti-sigma-factor
#11
Karlijn C Bastiaansen, Cristina Civantos, Wilbert Bitter, María A Llamas
Cell-surface signaling (CSS) is a signal transfer system that allows Gram-negative bacteria to detect environmental signals and generate a cytosolic response. These systems are composed of an outer membrane receptor that senses the inducing signal, an extracytoplasmic function sigma factor (σ(ECF)) that targets the cytosolic response by modifying gene expression and a cytoplasmic membrane anti-sigma factor that keeps the σ(ECF) in an inactive state in the absence of the signal and transduces its presence from the outer membrane to the cytosol...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28510364/lyophilization-of-bdellovibrio-bacteriovorus-109j-for-long-term-storage
#12
Mélanie J Boileau, Rinosh Mani, Kenneth D Clinkenbeard
Bdellovibrio bacteriovorus 109J is a Gram-negative predatory bacterium with obligate host dependency on other Gram-negative bacteria. This bacteriolytic predator collides with, enters, and establishes growth within the prey (host) periplasm, eventually lysing the prey cell wall to release fresh, motile B. bacteriovorus progeny. Laboratory maintenance of B. bacteriovorus has been previously described by other investigators. The protocols included in this unit deal with the technique required to lyophilize or freeze dry host-dependent B...
May 16, 2017: Current Protocols in Microbiology
https://www.readbyqxmd.com/read/28507245/conformational-changes-of-inter-domain-linker-mediate-mechanical-signal-transmission-in-sensor-kinase-bvgs
#13
Elodie Lesne, Elian Dupré, Camille Locht, Rudy Antoine, Françoise Jacob-Dubuisson
The whooping cough agent, Bordetella pertussis, controls the expression of its large virulence regulon in a coordinated manner through the two-component system BvgAS. BvgS is a dimeric, multi-domain sensor-kinase. Each monomer comprises in succession tandem periplasmic Venus flytrap (VFT) domains, a transmembrane segment, a cytoplasmic Per-ArnT-Sim (PAS) domain, a kinase module and additional phosphorelay domains. BvgS shifts between kinase and phosphatase modes of activity in response to chemical modulators that modify the clamshell motions of the VFT domains...
May 15, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28504659/structure-of-the-macab-tolc-abc-type-tripartite-multidrug-efflux-pump
#14
Anthony W P Fitzpatrick, Salomé Llabrés, Arthur Neuberger, James N Blaza, Xiao-Chen Bai, Ui Okada, Satoshi Murakami, Hendrik W van Veen, Ulrich Zachariae, Sjors H W Scheres, Ben F Luisi, Dijun Du
The MacA-MacB-TolC assembly of Escherichia coli is a transmembrane machine that spans the cell envelope and actively extrudes substrates, including macrolide antibiotics and polypeptide virulence factors. These transport processes are energized by the ATPase MacB, a member of the ATP-binding cassette (ABC) superfamily. We present an electron cryo-microscopy structure of the ABC-type tripartite assembly at near-atomic resolution. A hexamer of the periplasmic protein MacA bridges between a TolC trimer in the outer membrane and a MacB dimer in the inner membrane, generating a quaternary structure with a central channel for substrate translocation...
May 15, 2017: Nature Microbiology
https://www.readbyqxmd.com/read/28500346/structural-studies-of-the-periplasmic-portion-of-the-diguanylate-cyclase-cdgh-from-vibrio-cholerae
#15
Min Xu, Yi-Zhi Wang, Xiu-An Yang, Tao Jiang, Wei Xie
Cyclic diguanylate monophosphate (c-di-GMP) is a second messenger involved in bacterial signal transduction and produced by diguanylate cyclases (DGCs) generally containing highly variable periplasmic signal-recognition domains. CdgH is a DGC enzyme that regulates rugosity associated phenotypes in Vibrio cholerae. CdgH has two N-terminal tandem periplasmic substrate-binding (PBPb) domains for its signal recognition; however, the role of the tandem PBPb domains remains unclear. Here, we reported the crystal structure of the periplasmic portion of CdgH, which indicated that both tandem PBPb domains consist of typical interlobe ligand-binding architecture...
May 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28497288/novel-signal-peptides-improve-the-secretion-of-recombinant-staphylococcus-aureus-alpha-toxinh35l-in-escherichia-coli
#16
SooJin Han, Shushil Machhi, Mark Berge, Guoling Xi, Thomas Linke, Ronald Schoner
Secretion of heterologous proteins into Escherichia coli cell culture medium offers significant advantages for downstream processing over production as inclusion bodies; including cost and time savings, and reduction of endotoxin. Signal peptides play an important role in targeting proteins for translocation across the cytoplasmic membrane to the periplasmic space and release into culture medium during the secretion process. Alpha toxinH35L (ATH35L) was selected as an antigen for vaccine development against Staphylococcus aureus infections...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28492977/chaperone-substrate-interactions-monitored-via-a-robust-tem-1-%C3%AE-lactamase-fragment-complementation-assay
#17
Ling Bai, Wei He, Tianpeng Li, Cuiting Yang, Yingping Zhuang, Shu Quan
OBJECTIVE: To investigate the application of the TEM-1 β-lactamase protein fragment complementation assay (PCA) in detecting weak and unstable protein-protein interactions as typically observed during chaperone-assisted protein folding in the periplasm of Escherichia coli. RESULTS: The TEM-1 β-lactamase PCA system effectively captured the interactions of three pairs of chaperones and substrates. Moreover, the strength of the interactions can be quantitatively analyzed by comparing different levels of penicillin resistance, and the assay can be performed under 0...
May 10, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28488120/efficient-antibody-assembly-in-e-coli-periplasm-by-disulfide-bond-folding-factor-co-expression-and-culture-optimization
#18
Carlos Rodriguez, Dong Hyun Nam, Evan Kruchowy, Xin Ge
Molecular chaperones and protein folding factors of bacterial periplasmic space play important roles in assisting disulfide bond formation and proper protein folding. In this study, effects of disulfide bond protein (Dsb) families were investigated on assembly of 3F3 Fab, an antibody inhibitor targeting matrix metalloproteinase-14 (MMP-14). By optimizing DsbA/C co-expression, promoter for 3F3 Fab, host strains, and culture media and conditions, a high yield of 30-mg purified 3F3 Fab per liter culture was achieved...
May 10, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28487432/deciphering-mcr-2-colistin-resistance
#19
Jian Sun, Yongchang Xu, Rongsui Gao, Jingxia Lin, Wenhui Wei, Swaminath Srinivas, Defeng Li, Run-Shi Yang, Xing-Ping Li, Xiao-Ping Liao, Ya-Hong Liu, Youjun Feng
Antibiotic resistance is a prevalent problem in public health worldwide. In general, the carbapenem β-lactam antibiotics are considered a final resort against lethal infections by multidrug-resistant bacteria. Colistin is a cationic polypeptide antibiotic and acts as the last line of defense for treatment of carbapenem-resistant bacteria. Very recently, a new plasmid-borne colistin resistance gene, mcr-2, was revealed soon after the discovery of the paradigm gene mcr-1, which has disseminated globally. However, the molecular mechanisms for MCR-2 colistin resistance are poorly understood...
May 9, 2017: MBio
https://www.readbyqxmd.com/read/28487352/in-vivo-structures-of-an-intact-type-vi-secretion-system-revealed-by-electron-cryotomography
#20
Yi-Wei Chang, Lee A Rettberg, Davi R Ortega, Grant J Jensen
The type VI secretion system (T6SS) is a versatile molecular weapon used by many bacteria against eukaryotic hosts or prokaryotic competitors. It consists of a cytoplasmic bacteriophage tail-like structure anchored in the bacterial cell envelope via a cytoplasmic baseplate and a periplasmic membrane complex. Rapid contraction of the sheath in the bacteriophage tail-like structure propels an inner tube/spike complex through the target cell envelope to deliver effectors. While structures of purified contracted sheath and purified membrane complex have been solved, because sheaths contract upon cell lysis and purification, no structure is available for the extended sheath...
May 9, 2017: EMBO Reports
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