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outer membrane protein analysis

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https://www.readbyqxmd.com/read/29355812/lack-of-cations-in-flow-cytometry-buffers-affect-fluorescence-signals-by-reducing-membrane-stability-and-viability-of-escherichia-coli-strains
#1
Kathrin Tomasek, Tobias Bergmiller, Călin C Guet
Buffers are essential for diluting bacterial cultures for flow cytometry analysis in order to study bacterial physiology and gene expression parameters based on fluorescence signals. Using a variety of constitutively expressed fluorescent proteins in Escherichia coli K-12 strain MG1655, we found strong artifactual changes in fluorescence levels after dilution into the commonly used flow cytometry buffer phosphate-buffered saline (PBS) and two other buffer solutions, Tris-HCl and M9 salts. These changes appeared very rapidly after dilution, and were linked to increased membrane permeability and loss in cell viability...
January 17, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29345850/effects-of-glutaraldehyde-didecyldimethylammonium-bromide-combined-disinfectant-on-the-cell-surface-of-staphylococcus-aureus
#2
Wenshu Lin, Jialin Yi, Yuelin Zhang, Zhirui Deng, Qin Chen, Bing Niu
AIMS: The effects of a new glutaraldehyde-didecyldimethylammonium bromide combination disinfectant (GD) on the cell surface of Staphylococcus aureus, a representative Gram-positive bacterium, were investigated in this study. METHODS AND RESULTS: Results of bacterial surface structural analysis showed that GD significantly changed the bacterial morphology. The membrane fluidity decreased and outer membrane permeabilization increased after contact with GD. Furthermore, the integrity of the cytoplasmic membrane was destroyed by over 99% after exposure to GD for a short time...
January 18, 2018: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29339308/antibiotic-resistance-and-virulence-traits-of-bacterial-pathogens-from-infected-freshwater-fish-labeo-rohita
#3
Dharmaraj Ramesh, Sami Souissi
Bacterial infectious diseases are a main dangerous problem in Aquaculture farming. It causes multiple diseases in fish as well as in human being and it has considerable virulence potential. In this connection, the moot of study focus to discriminate bacterial isolates recovered from naturally diseased Labeo rohita fish and their virulent characteristics. Based on the β-haemolysis factor, four isolates (KADR11, KADR12, KADR13 and KADR14) were selected for further delineation. These bacterial isolates showed high similarity with Providencia rettgeri, Aeromonas sp...
January 12, 2018: Microbial Pathogenesis
https://www.readbyqxmd.com/read/29327006/spectrochemical-analyses-of-growth-phase-related-bacterial-responses-to-low-environmentally-relevant-concentrations-of-tetracycline-and-nanoparticulate-silver
#4
Naifu Jin, Kirk T Semple, Longfei Jiang, Chunling Luo, Dayi Zhang, Francis L Martin
Exposure to environmental insults generally occurs at low levels, making it challenging to measure bacterial responses to such interactions. Additionally, microbial behaviour and phenotype varies in differing bacterial types or growth phases, likely giving rise to growth- or species-specific responses to environmental stimuli. The present study applied a spectrochemical tool, infrared (IR) spectral interrogation coupled with multivariate analysis, to investigate the growth- and species-specific responses of two bacterial strains, Gram-negative Pseudomonas fluorescens and Gram-positive Mycobacterium vanbaalenii, to low concentrations of tetracycline, nanoparticulate silver (AgNP) or mixtures thereof...
January 12, 2018: Analyst
https://www.readbyqxmd.com/read/29326362/-18f-pbr111-pet-imaging-in-healthy-controls-and-schizophrenia-test-retest-reproducibility-and-quantification-of-neuroinflammation
#5
Julie Ottoy, Livia De Picker, Jeroen Verhaeghe, Steven Deleye, Leonie Wyffels, Lauren Kosten, Bernard Sabbe, Violette Coppens, Maarten Timmers, Luc Van Nueten, Sarah Ceyssens, Sigrid Stroobants, Manuel Morrens, Steven Staelens
Activated microglia express the translocator protein (TSPO) on the outer mitochondrial membrane. 18F-PBR111 is a second-generation positron emission tomography (PET) ligand that specifically binds the TSPO, allowing in-vivo visualization and quantification of neuroinflammation. The aim of this study is to evaluate if the test-retest variability of 18F-PBR111 in healthy controls is acceptable to detect a psychosis-associated neuroinflammatory signal in schizophrenia. Methods: Dynamic 90-min 18F-PBR111 scans were obtained in 17 healthy male controls (HC) and 11 male schizophrenia patients during a psychotic episode (SP)...
January 11, 2018: Journal of Nuclear Medicine: Official Publication, Society of Nuclear Medicine
https://www.readbyqxmd.com/read/29324785/phylogenetic-diversity-antimicrobial-susceptibility-and-virulence-gene-profiles-of-brachyspira-hyodysenteriae-isolates-from-pigs-in-germany
#6
Jessica Joerling, Stefanie A Barth, Karen Schlez, Hermann Willems, Werner Herbst, Christa Ewers
Swine dysentery (SD) is an economically important diarrheal disease in pigs caused by different strongly hemolytic Brachyspira (B.) species, such as B. hyodysenteriae, B. suanatina and B. hampsonii. Possible associations of epidemiologic data, such as multilocus sequence types (STs) to virulence gene profiles and antimicrobial susceptibility are rather scarce, particularly for B. hyodysenteriae isolates from Germany. In this study, B. hyodysenteriae (n = 116) isolated from diarrheic pigs between 1990 and 2016 in Germany were investigated for their STs, susceptibility to the major drugs used for treatment of SD (tiamulin and valnemulin) and genes that were previously linked with virulence and encode for hemolysins (tlyA, tlyB, tlyC, hlyA, BHWA1_RS02885, BHWA1_RS09085, BHWA1_RS04705, and BHWA1_RS02195), outer membrane proteins (OMPs) (bhlp16, bhlp17...
2018: PloS One
https://www.readbyqxmd.com/read/29324691/physiological-and-comparative-genomic-analysis-of-arthrobacter-sp-srs-w-1-2016-provides-insights-on-niche-adaptation-for-survival-in-uraniferous-soils
#7
Ashvini Chauhan, Ashish Pathak, Rajneesh Jaswal, Bobby Edwards Iii, Demario Chappell, Christopher Ball, Reyna Garcia-Sillas, Paul Stothard, John Seaman
Arthrobacter sp. strain SRS-W-1-2016 was isolated on high concentrations of uranium (U) from the Savannah River Site (SRS) that remains co-contaminated by radionuclides, heavy metals, and organics. SRS is located on the northeast bank of the Savannah River (South Carolina, USA), which is a U.S. Department of Energy (DOE) managed ecosystem left historically contaminated from decades of nuclear weapons production activities. Predominant contaminants within the impacted SRS environment include U and Nickel (Ni), both of which can be transformed microbially into less toxic forms via metal complexation mechanisms...
January 11, 2018: Genes
https://www.readbyqxmd.com/read/29315985/comparative-analysis-of-the-hom-family-of-outer-membrane-proteins-in-isolates-from-two-geographically-distinct-regions-the-united-states-and-south-korea
#8
Stephanie L Servetas, Aeryun Kim, Hanfu Su, Jeong-Heon Cha, D Scott Merrell
BACKGROUND: Helicobacter pylori encodes numerous outer membrane proteins (OMPs), but only a few have been characterized in depth. Deletion, duplication, and allelic variation of many of the H. pylori OMPs have been reported, which suggests that these proteins may play key roles in host adaptation. Herein, we characterize the variation observed within the Hom family of OMPs in H. pylori obtained from two geographically distinct populations. MATERIALS AND METHODS: PCR genotyping of the hom genes was carried out using clinical isolates from South Korea and the United States...
January 5, 2018: Helicobacter
https://www.readbyqxmd.com/read/29311259/the-endosomal-trafficking-factors-corvet-and-escrt-suppress-plasma-membrane-residence-of-the-renal-outer-medullary-potassium-channel-romk
#9
Timothy D Mackie, Bo-Young Kim, Arohan R Subramanya, Daniel J Bain, Allyson F O'Donnell, Paul A Welling, Jeffrey L Brodsky
Protein trafficking can act as the primary regulatory mechanism for ion channels with high open probabilities, such as the renal outer medullary (ROMK) channel. ROMK, also known as Kir1.1 (KCNJ1), is the major route for potassium secretion into the pro-urine and plays an indispensable role in regulating serum potassium and urinary concentrations. However, the cellular machinery that regulates ROMK trafficking has not been fully defined. To identify regulators of the cell-surface population of ROMK, we expressed a pH-insensitive version of the channel in the budding yeast Saccharomyces cerevisiae...
January 8, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29299929/proteome-of-bovine-mitochondria-and-rod-outer-segment-disks-commonalities-and-differences
#10
Maurizio Bruschi, Andrea Petretto, Federico Caicci, Martina Bartolucci, Daniela Calzia, Laura Santucci, Lucia Manni, Luca A Ramenghi, Gian Marco Ghiggeri, Carlo E Traverso, Giovanni Candiano, Isabella Panfoli
The retinal rod outer segment (OS) is a stack of disks surrounded by the plasma membrane, housing proteins related to phototransduction, as well as mitochondrial proteins involved in oxidative phosphorylation (OxPhos). This prompted us to compare the proteome of bovine OS disks and mitochondria, to assess the significant top gene signatures of each sample. The two proteomes, obtained by LTQ-Orbitrap Velos mass spectrometry, were compared by statistical analyses. 4139 proteins were identified, 2045 of which overlapping in the two sets...
January 4, 2018: Journal of Proteome Research
https://www.readbyqxmd.com/read/29298884/genomic-and-biochemical-characterization-of-acinetobacter-podophage-petty-reveals-a-novel-lysis-mechanism-and-tail-associated-depolymerase-activity
#11
A C Hernandez-Morales, L L Lessor, T L Wood, D Migl, E M Mijalis, W K Russell, R F Young, J J Gill
The increased prevalence of drug-resistant, nosocomial Acinetobacter infections, particularly from pathogenic members of the Acinetobacter calcoaceticus-baumannii complex, necessitates the exploration of novel treatments such as phage therapy. In the present study, we characterize phage Petty, a novel podophage that infects multidrug-resistant Acinetobacter nosocomialis and Acinetobacter baumannii Genome analysis reveals that phage Petty is a 40,431bp ϕKMV-like phage, with a coding density of 92.2% and a G+C content of 42...
January 3, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29297821/isolation-identification-and-genome-analysis-of-lung-pathogenic-klebsiella-pneumoniae-lpkp-in-forest-musk-deer
#12
Wei Zhao, Qing Tian, Yan Luo, Yin Wang, Ze-Xiao Yang, Xue-Ping Yao, Jian-Guo Cheng, Xin Zhou, Wu-You Wang
Klebsiella pneumoniae is an important pathogen commonly associated with opportunistic infections. In this study, lung pathogenic K. pneumoniae (LPKP) was isolated and identified from suppurative pneumoniae in forest musk deer by conventional methods and by 16S ribosomal RNA sequence analysis. Median lethal dose and histopathologic analysis were used to demonstrate pathogenicity of the organism in mice. Furthermore, a draft genome of LPKP was sequenced, and its virulence genes were detected. One hundred and twenty-two virulence genes encoded determinant of capsule polysaccharide (CPS), lipopolysaccharide, fimbriae, outer membrane proteins, iron acquisition, and urease...
December 2017: Journal of Zoo and Wildlife Medicine: Official Publication of the American Association of Zoo Veterinarians
https://www.readbyqxmd.com/read/29296265/molecular-characterization-and-functional-analysis-of-the-pilq380-706-a-novel-secretin-domain-in-pseudomonas-aeruginosa
#13
Sobhan Faezi, Iraj Nikokar, Ali Elmi, Yusuf Ghasemi, Mojtaba Farahbakhsh, Alireza Salimi Chirani, Mehdi Mahdavi
Background: Type 4 pili (T4P) is an important virulence factor of Pseudomonas aeruginosa (P. aeruginosa). T4P pass the outer membrane through a large oligomeric channel made of a single PilQ protein that is most highly conserved at their C-termini. To develop a functional vaccine that can be used in clinical application, the secretin domain of the PilQ (PilQ380-706) was produced as a recombinant protein. Methods: A 981 bp fragment of C-terminal of the pilQ secretin (pilQ1138-2118) from was designed into the prokaryotic expression vector pET28a...
January 2018: Avicenna Journal of Medical Biotechnology
https://www.readbyqxmd.com/read/29291378/a-new-use-of-%C3%AE-ala-lys-amca-as-a-transport-reporter-for-pept1-and-pept2-in-renal-brush-border-membrane-vesicles-from-the-outer-cortex-and-outer-medulla
#14
Othman A Alghamdi, Nicola King, Graham L Jones, Pierre D J Moens
Integral membrane proteins PEPT1 and PEPT2 are essential for reabsorbing almost all hydrolysed or filtered di- and tripeptides alongside a wide range of peptidomimetic drugs in the kidney. The aim of this study was to investigate the potential use of the fluorophore-conjugated dipeptide β-Ala-Lys (AMCA) as a biosensor for measuring peptide transport activity in brush border membrane vesicles isolated from the outer cortex (BBMV-OC) and outer medulla (BBMV-OM) (representing PEPT1 and PEPT2 respectively). The vesicles were isolated using a dual magnesium precipitation and centrifugation technique...
December 29, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29290623/identification-of-new-activators-of-mitochondrial-fusion-reveals-a-link-between-mitochondrial-morphology-and-pyrimidine-metabolism
#15
Laia Miret-Casals, David Sebastián, José Brea, Eva M Rico-Leo, Manuel Palacín, Pedro M Fernández-Salguero, M Isabel Loza, Fernando Albericio, Antonio Zorzano
Mitochondria are dynamic organelles that produce most of the cellular ATP, and are involved in many other cellular functions such as Ca2+ signaling, differentiation, apoptosis, cell cycle, and cell growth. One key process of mitochondrial dynamics is mitochondrial fusion, which is catalyzed by mitofusins (MFN1 and MFN2) and OPA1. The outer mitochondrial membrane protein MFN2 plays a relevant role in the maintenance of mitochondrial metabolism, insulin signaling, and mutations that cause neurodegenerative disorders...
December 23, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/29289381/recombinant-lactococcus-lactis-co-expressing-omph-of-an-m-cell-targeting-ligand-and-ibdv-vp2-protein-provide-immunological-protection-in-chickens
#16
Linlin Liu, Wang Zhang, Yuxin Song, Wenqian Wang, Yuan Zhang, Tingting Wang, Kai Li, Qing Pan, Xiaole Qi, Yulong Gao, Li Gao, Changjun Liu, Yanping Zhang, Yongqiang Wang, Gaoming He, Xiaomei Wang, Hongyu Cui
Infectious bursal disease virus (IBDV) is a highly contagious disease that results in enormous economic losses in the global poultry sector. Lactic acid bacteria are an appealing vehicle for the safe and effective delivery of heterologous protein antigens. Oral administration of the commensal bacterium Lactococcus lactis expressing recombinant fusion proteins has been used to elicit mucosal and systemic immune responses. In this study, a Lactococcus lactis NZ3900 strain co-expressing the outer membrane protein (Omp) H of the microfold (M) cell-targeting ligand and the viral capsid protein (VP)2 antigen of IBDV was genetically engineered, and its immunopotentiating capacity as an oral and injected vaccine in chickens was evaluated...
December 27, 2017: Vaccine
https://www.readbyqxmd.com/read/29288627/characterization-of-the-first-planctomycetal-outer-membrane-protein-identifies-a-channel-in-the-outer-membrane-of-the-anammox-bacterium-kuenenia-stuttgartiensis
#17
Muriel C F van Teeseling, Roland Benz, Naomi M de Almeida, Mike S M Jetten, Rob J Mesman, Laura van Niftrik
Planctomycetes are a bacterial phylum known for their complex intracellular compartmentalization. While most Planctomycetes have two compartments, the anaerobic ammonium oxidizing (anammox) bacteria contain three membrane-enclosed compartments. In contrast to a long-standing consensus, recent insights suggested the outermost Planctomycete membrane to be similar to a Gram-negative outer membrane (OM). One characteristic component that differentiates OMs from cytoplasmic membranes (CMs) is the presence of outer membrane proteins (OMPs) featuring a ß-barrel structure that facilitates passage of molecules through the OM...
December 27, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29284141/optimization-of-haemophilus-influenzae-adhesin-transmembrane-domain-expression-in-escherichia-coli
#18
Eriko Aoki, Kazuo Fujiwara, Akio Shimizu, Sayaka Takase-Yoden, Masamichi Ikeguchi
To obtain a high yield of the transmembrane domain of Haemophilus influenzae adhesin (HiaTD) in Escherichia coli, we attempted to express the HiaTD with and without a signal sequence using a T7 expression system. The expression level of HiaTD after induction was followed by quantification of the purified HiaTD, flow cytometric analysis of the outer membrane integrated HiaTD, and immunoblotting assay of fractionated cell lysate. In the expression system with a signal sequence, although the amount of cell-surface-expressed HiaTD increased over time, the number of HiaTD-expressing cells decreased, probably because of plasmid instability...
December 25, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/29284024/diabetes-related-changes-in-the-protein-composition-and-the-biomechanical-properties-of-human-retinal-vascular-basement-membranes
#19
Willi Halfter, Suzette Moes, Daphne O Asgeirsson, Kathrin Halfter, Philipp Oertle, Esther Melo Herraiz, Marija Plodinec, Paul Jenoe, Paul Bernhard Henrich
Basement membranes (BMs) are specialized sheets of extracellular matrix that outline epithelial cell layers, muscle fibers, blood vessels, and peripheral nerves. A well-documented histological hallmark of progressing diabetes is a major increase in vascular BM thickness. In order to investigate whether this structural change is accompanied by a change in the protein composition, we compared the proteomes of retinal vascular BMs from diabetic and non-diabetic donors by using LC-MS/MS. Data analysis showed that seventeen extracellular matrix (ECM)-associated proteins were more abundant in diabetic than non-diabetic vascular BMs...
2017: PloS One
https://www.readbyqxmd.com/read/29280343/the-ropae-gene-encodes-a-porin-like-protein-involved-in-copper-transit-in-rhizobium-etli-cfn42
#20
Antonio González-Sánchez, Ciro A Cubillas, Fabiola Miranda, Araceli Dávalos, Alejandro García-de Los Santos
Copper (Cu) is an essential micronutrient for all aerobic forms of life. Its oxidation states (Cu+ /Cu2+ ) make this metal an important cofactor of enzymes catalyzing redox reactions in essential biological processes. In gram-negative bacteria, Cu uptake is an unexplored component of a finely regulated trafficking network, mediated by protein-protein interactions that deliver Cu to target proteins and efflux surplus metal to avoid toxicity. Rhizobium etliCFN42 is a facultative symbiotic diazotroph that must ensure its appropriate Cu supply for living either free in the soil or as an intracellular symbiont of leguminous plants...
December 27, 2017: MicrobiologyOpen
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