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outer membrane protein analysis

Khalid Shahin, Kim D Thompson, Neil F Inglis, Kevin Mclean, Jose Gustavo Ramirez-Paredes, Sean J Monaghan, Rowena Hoare, Michael Fontaine, Matthijs Metselaar, Alexandra Adams
AIMS: The aims of the current study were to characterise the outer membrane proteins (OMPs) of Francisella noatunensis subsp. orientalis (Fno) STIR-GUS-F2f7, and identify proteins recognised by sera from tilapia, Oreochromis niloticus, (L) that survived experimental challenge with Fno. METHODS AND RESULTS: The composition of the OMPs of a virulent strain of Fno (STIR-GUS- F2f7), isolated from diseased red Nile tilapia in UK, was examined. The sarcosine-insoluble OMP fraction was screened with tilapia hyper-immune sera by western blot analysis following separation of the proteins by 1D SDS-PAGE...
May 19, 2018: Journal of Applied Microbiology
Sultana Rasheed, Khurram Bashir, Jong-Myong Kim, Marina Ando, Maho Tanaka, Motoaki Seki
The Arabidopsis histone deacetylase 6 (HDA6) mutant exhibits increased tolerance to drought stress by negatively regulating the expression of ALDH2B7 and PDC1. Therefore, it was logical to determine if transgenic Arabidopsis plants expressing PDC1 or ALDH2B7 using a suitable promoter would also exhibit tolerance to drought stress. An analysis of published microarray data indicated the up-regulation of the TSPO gene, which encodes an outer membrane tryptophan-rich sensory protein (TSPO), by drought stress. RT-qPCR, as well as GUS analysis of the promoter, confirmed the up-regulation of TSPO by drought stress in Arabidopsis roots and shoots...
May 18, 2018: Scientific Reports
Min Woo Sung, Rahamthulla Shaik, Allan D TerBush, Katherine W Osteryoung, Stanislav Vitha, Andreas Holzenburg
Chloroplasts host photosynthesis and fulfill other metabolic functions that are essential to plant life. They have to divide by binary fission to maintain their numbers throughout cycles of cell division. Chloroplast division is achieved by a complex ring-shaped division machinery located on both the inner (stromal) and the outer (cytosolic) side of the chloroplast envelope. The inner division ring (termed the Z ring) is formed by the assembly of tubulin-like FtsZ1 and FtsZ2 proteins. ARC6 is a key chloroplast division protein that interacts with the Z ring...
May 16, 2018: Journal of Biological Chemistry
Joseph J Wanford, Luke R Green, Jack Aidley, Christopher D Bayliss
Pathogenic Neisseria are responsible for significantly higher levels of morbidity and mortality than their commensal relatives despite having similar genetic contents. Neisseria possess a disparate arsenal of surface determinants that facilitate host colonisation and evasion of the immune response during persistent carriage. Adaptation to rapid changes in these hostile host environments is enabled by phase variation (PV) involving high frequency, stochastic switches in expression of surface determinants. In this study, we analysed 89 complete and 79 partial genomes, from the NCBI and Neisseria PubMLST databases, representative of multiple pathogenic and commensal species of Neisseria using PhasomeIt, a new program that identifies putatively phase-variable genes and homology groups by the presence of simple sequence repeats (SSR)...
2018: PloS One
Priya Uppuluri, Lin Lin, Abdullah Alqarihi, Guanpingsheng Luo, Eman G Youssef, Sondus Alkhazraji, Nannette Y Yount, Belal A Ibrahim, Michael Anthony Bolaris, John E Edwards, Marc Swidergall, Scott G Filler, Michael R Yeaman, Ashraf S Ibrahim
Different pathogens share similar medical settings and rely on similar virulence strategies to cause infections. We have previously applied 3-D computational modeling and bioinformatics to discover novel antigens that target more than one human pathogen. Active and passive immunization with the recombinant N-terminus of Candida albicans Hyr1 (rHyr1p-N) protect mice against lethal candidemia. Here we determine that Hyr1p shares homology with cell surface proteins of the multidrug resistant Gram negative bacterium, Acinetobacter baumannii including hemagglutinin (FhaB) and outer membrane protein A (OmpA)...
May 10, 2018: PLoS Pathogens
Stephan Klatt, Rajini Brammananth, Sean O'Callaghan, Konstantinos Andreas Kouremenos, Dedreia Tull, Paul K Crellin, Ross L Coppel, Malcolm J McConville
The complex cell envelopes of Corynebacterineae contribute to the virulence of pathogenic species (such as Mycobacterium tuberculosis and Corynebacterium diphtheriae) and capacity of non-pathogenic species (such as Corynebacterium glutamicum) to grow in diverse niches. The Corynebacterineae cell envelope comprises an asymmetric outer membrane, that overlays the arabinogalactan-peptidoglycan complex and the inner cell membrane. Dissection of the lipid composition of the inner and outer membrane fractions is important for understanding the biogenesis of this multi-laminate wall structure...
May 3, 2018: Journal of Lipid Research
Yan-Jiao Zhang, Huiyuan Lin, Pan Wang, Chang Chen, Shiyong Chen
Vibrio vulnificus, a Gram-negative halophilic bacterium, is an opportunistic human pathogen that is responsible for the majority of seafood-associated deaths worldwide. Lipoproteins are important components of the bacterial cell envelope and have been shown to be involved in a wide variety of cellular processes. Little is known about the localisation or transport mechanism of lipoproteins in V. vulnificus. To assess the localisation of lipoproteins in V. vulnificus, we tested two established techniques for the rapid separation of membrane-associated proteins: detergent extraction with Sarkosyl and outer membrane vesicles (OMVs) preparation...
May 2, 2018: Antonie Van Leeuwenhoek
Young Bin Im, Woo Bin Park, Myunghwan Jung, Suk Kim, Han Sang Yoo
Upon infection by Brucella, cytokine production is induced which becomes an important factor in evaluation of innate and adaptive immune responses. Several research groups have been investigating the mechanisms involved in Brucella infection within the host. Hence, we conducted an analytical study to define pathogenic pathways and immune mechanisms upon Brucella infection by investigating the efficacy of recombinant outer membrane protein 10 (rOMP10), outer membrane protein 19 (rOMP19) and outer membrane protein 28 (rOMP28) in in vitro and in vivo upon stimulation/immunization...
April 27, 2018: Japanese Journal of Infectious Diseases
Tsitsi D Mubaiwa, Lauren E Hartley-Tassell, Evgeny A Semchenko, Christopher J Day, Michael P Jennings, Kate L Seib
Neisseria meningitidis is a Gram-negative bacterial pathogen that causes life threatening meningitis and septicemia. Neisseria Heparin Binding Antigen (NHBA) is an outer membrane protein that binds heparin and heparan sulfate and DNA. This protein is one of the four antigens in the meningococcal serogroup B vaccine Bexsero. In the current study, we sought to define the full glycan-binding repertoire of NHBA to better understand its role in meningococcal pathogenesis and vaccine efficacy. Glycan array analysis revealed binding to 28 structures by recombinant NHBA...
April 25, 2018: Scientific Reports
Xiaojie Song, Hua Zhang, Dongsheng Zhang, Weifeng Xie, Guanghui Zhao
The clinical isolation rate of multidrug-resistant or pan-resistant Acinetobacter baumannii (A. baumannii) is increasing, resulting that optional antibiotics are very limited in clinical practice. To deal with such a dilemma in treatment, the development of effective vaccines serves as a good strategy. Outer membrane proteins (Omp) often contain potential excellent vaccine antigens, and NCBI has published >300 Omp sequences of A. baumannii (including the duplicates). To accurately screen out the potential excellent antigen molecules from a large number of sequences, and avoid repetitive experimental processes is of great significance...
April 16, 2018: Infection, Genetics and Evolution
Rui Qing, Qingke Xiang, Zhongqi Liu, Fei Xiao, Fan Yang
Objective To investigate the effect of aluminum phosphate (AP) and aluminum hydroxide (AH) as adjuvants on Brucella outer membrane protein 31 (Omp31) in inducing humoral and cellular immune responses and immune protection. Methods AP and AH adjuvants were prepared and separately mixed with Brucella Omp31 protein to measure the adsorption rates. The AP- and AH-absorbed Omp31 protein were intraperitoneally injected into BLAB/c mice at 0, 2, and 4 weeks, and meanwhile, unabsorbed Omp31 protein and PBS were used as controls...
February 2018: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Rodolfo Villena, Marco Aurelio P Safadi, María Teresa Valenzuela, Juan P Torres, Adam Finn, Miguel O'Ryan
Meningococcal disease (MD) is a major cause of meningitis and sepsis worldwide, with a high case fatality rate and frequent sequelae. Neisseria meningitidis serogroups A, B, C, W, X and Y are responsible for most of these life-threatening infections, and its unpredictable epidemiology can cause outbreaks in communities, with significant health, social and economic impact. Currently, serogroup B is the main cause of MD in Europe and North America and one of the most prevalent serogroups in Latin America. Mass vaccination strategies using polysaccharide vaccines have been deployed since the 1970s and the use of conjugate vaccines has controlled endemic and epidemic disease caused by serogroups A, C, W and Y and more recently serogroup B using geographically-specific outer membrane vesicle based vaccines...
April 18, 2018: Human Vaccines & Immunotherapeutics
Saori Maki-Yonekura, Rei Matsuoka, Yoshiki Yamashita, Hirofumi Shimizu, Maiko Tanaka, Fumie Iwabuki, Koji Yonekura
Gram-negative bacteria import essential nutrients such as iron and vitamin B12 through outer membrane receptors. This process utilizes proton motive force harvested by the Ton system made up of three inner membrane proteins, ExbB, ExbD and TonB. ExbB and ExbD form the proton channel that energizes uptake through TonB. Recently, crystal structures suggest that the ExbB pentamer is the scaffold. Here, we present structures of hexameric complexes of ExbB and ExbD revealed by X-ray crystallography and single particle cryo-EM...
April 17, 2018: ELife
Mariana Papalia, Germán Traglia, Melina Ruggiero, Marisa Almuzara, Carlos Vay, Gabriel Gutkind, Soledad María Ramírez, Marcela Radice
OBJECTIVES: The aim of this study was to characterize OXA-258 variants and other features that may contribute to the carbapenem resistant in Achromobacter ruhlandii. METHODS: Kinetic parameters for purified OXA-258a and OXA-258b were determined measuring the rate of hydrolysis of a representative group of antibiotics. Whole-genome shotgun sequencing was performed on A. ruhlandii 38 (producing OXA-258a) and A. ruhlandii 319 (producing OXA-258b) and in silico analysis of antimicrobial resistant determinants was conducted...
April 9, 2018: Journal of Global Antimicrobial Resistance
Xiaojun Yuan, Matthew D Johnson, Jing Zhang, Alvin W Lo, Mark A Schembri, Lakshmi C Wijeyewickrema, Robert N Pike, Gerard H M Huysmans, Ian R Henderson, Denisse L Leyton
Bacterial autotransporters comprise a C-terminal β-barrel domain, which must be correctly folded and inserted into the outer membrane to facilitate translocation of the N-terminal passenger domain to the cell exterior. Once at the surface, the passenger domains of most autotransporters are folded into an elongated β-helix. In a cellular context, key molecules catalyze the assembly of the autotransporter β-barrel domain. However, how the passenger domain folds into its functional form is poorly understood...
April 11, 2018: Nature Communications
Qian Wang, Shanshan Zhao, Hazi Wureli, Songsong Xie, Chuangfu Chen, Qingqing Wei, Buyun Cui, Changchun Tu, Yuanzhi Wang
Brucellosis is a relatively common zoonosis, particularly in the developing countries. Our aim was to cultivate the Brucella wild-type strain from eggs of Dermacentor marginatus, and to molecularly investigate the presence of Brucella DNA in different developmental stages. A total of 350 engorged adult female ticks were collected from sheep and cattle, and individually placed into a single clean breathable tube suitable for laying eggs and for the development of larvae. Detection of Brucella DNA was attempted in engorged female ticks and their offsprings based on the Brucella outer membrane protein gene 22 (omp22) and IS711 gene...
March 26, 2018: Ticks and Tick-borne Diseases
Danilo Donnarumma, Claudio Maestri, Pietro Ivan Giammarinaro, Luigi Capriotti, Erika Bartolini, Daniele Veggi, Roberto Petracca, Maria Scarselli, Nathalie Norais
Hydrogen Deuterium exchange (HDx) associated with Mass Spectrometry (MS) is emerging as a powerful tool to provide conformational information on membrane proteins. Unfortunately, as for X-ray diffraction and NMR, HDx performed on reconstituted in vitro systems might not always reflect the in vivo environment. Outer Membrane Vesicles naturally released by E. coli were used to carry out analysis of native OmpF through HDx-MS. A new protocol compatible with HDx analysis and avoiding hindrance from the lipid contents was set-up...
April 5, 2018: Journal of Proteome Research
Leonardo A Guevarra, Andrea Claudine F Afable, Patricia Joyce O Belza, Karen Joy S Dy, Scott Justin Q Lee, Teresa T Sy-Ortin, Pia Marie S P Albano
Background: The role of Fusobacterium nucleatum Fap2 protein in the development of colorectal cancer has recently been explained. Fap2, when bound to the human inhibitory receptor, TIGIT, inhibits the cytotoxic activity of natural killer (NK) cells against cancer cells, thus, allowing proliferation of the latter eventually leading to tumor growth. The aim of the study was to identify the immunogenicity of a peptide mimotope of the Fap2 protein and to determine the reactivity of colorectal cancer patients' sera against the mimotope...
2018: Infectious Agents and Cancer
Kan-Na Tomioku, Mikiko Shigekuni, Hiroki Hayashi, Akane Yoshida, Taiki Futagami, Hisanori Tamaki, Kenji Tanabe, Akikazu Fujita
In budding yeast Saccharomyces cerevisiae, PtdIns(4)P serves as an essential signalling molecule in the Golgi complex, endosomal system, and plasma membrane, where it is involved in the control of multiple cellular functions via direct interactions with PtdIns(4)P-binding proteins. To analyse the distribution of PtdIns(4)P in yeast cells at a nanoscale level, we employed an electron microscopy technique that specifically labels PtdIns(4)P on the freeze-fracture replica of the yeast membrane. This method minimizes the possibility of artificial perturbation, because molecules in the membrane are physically immobilised in situ...
March 27, 2018: European Journal of Cell Biology
Si Won Kim, Seong Bin Park, Se Pyeong Im, Jung Seok Lee, Jae Wook Jung, Tae Won Gong, Jassy Mary S Lazarte, Jaesung Kim, Jong-Su Seo, Jong-Hwan Kim, Jong-Wook Song, Hyun Suk Jung, Gwang Joong Kim, Young Ju Lee, Suk-Kyung Lim, Tae Sung Jung
Outer membrane vesicles (OMVs) containing various bacterial compounds are released from mainly gram-negative bacteria. Secreted OMVs play important roles in the ability of a bacterium to defend itself, and thus contribute to the survival of bacteria in a community. In this study, we collected OMVs from β-lactam antibiotic-resistant Escherichia coli established by conjugation assay and the parental β-lactam antibiotic-susceptible strain, and performed comparative proteomic analysis to examine whether these OMVs carried β-lactam-resistant compounds...
March 29, 2018: Scientific Reports
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