Comparison of Extracellular Proteins from Virulent and Avirulent Vibrio parahaemolyticus Strains to Identify Potential Virulence Factors

2019 ◽  
Vol 83 (1) ◽  
pp. 155-162
Author(s):  
YU HE ◽  
SHUAI WANG ◽  
XIANTING YIN ◽  
FENGJIAO SUN ◽  
BIN HE ◽  
...  
Keyword(s):  
Foodborne Pathogens ◽  
Vibrio Parahaemolyticus ◽  
Stress Protein ◽  
Extracellular Proteins ◽  
Trigger Factor ◽  
Uncharacterized Protein ◽  
Adverse Conditions ◽  
Semialdehyde Dehydrogenase ◽  
Associated Proteins ◽  
Related Proteins

ABSTRACT Vibrio parahaemolyticus is a leading seafood-borne pathogen that causes gastroenteritis, septicemia, and serious wound infections due to the actions of virulence-associated proteins. We compared the extracellular proteins of nonvirulent JHY20 and virulent ATCC 33847 V. parahaemolyticus reference strains. Eighteen extracellular proteins were identified from secretory profiles, and 11 (68.75%) of the 16 proteins in ATCC 33847 are associated with virulence and/or protection against adverse conditions: trigger factor, chaperone SurA, aspartate–semialdehyde dehydrogenase, 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase, glutamate 5-kinase, alanine dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, outer membrane protein OmpV, ribosome-associated inhibitor A, chaperone protein Skp, and universal stress protein. Two nontoxic-related proteins, amino acid ABC transporter substrate-binding protein and an uncharacterized protein, were identified in JHY20. The results provide a theoretical basis for supporting safety risk assessment of aquatic foods, illuminate the pathogenic mechanisms of V. parahaemolyticus, and assist the identification of novel vaccine candidates for foodborne pathogens. HIGHLIGHTS

Comparative secretome analyses of toxigenic and atoxigenic Rathayibacter species

2021 ◽  
Author(s):  
Matthew Tancos ◽  
Michael McMahon ◽  
Wesley Garrett ◽  
Douglas G Luster ◽  
Elizabeth Rogers
Keyword(s):  
Plant Pathogen ◽  
Plant Pathogens ◽  
Extracellular Proteins ◽  
Department Of Agriculture ◽  
Homologous Proteins ◽  
Diagnostic Assays ◽  
Animal Pathogens ◽  
Associated Proteins ◽  
Inspection Service ◽  
Related Proteins

Phytopathogenic Rathayibacter species are unique bacterial plant pathogens as they are obligately vectored by plant parasitic Anguinid nematodes to the developing seedheads of forage grasses and cereals. This understudied group of plant-associated Actinomycetes includes the neurotoxigenic plant pathogen, R. toxicus, which causes annual ryegrass toxicity in grazing livestock. Rathayibacter toxicus is currently endemic to Australia and is listed as a plant pathogen select agent by the U.S. Department of Agriculture – Animal and Plant Health Inspection Service. The complex Rathayibacter disease cycle requires intimate interactions with the nematode vector and plant hosts, which warrants an increased understanding of the secretory and surface-associated proteins that mediate these diverse eukaryotic interactions. Here we present the first comparative secretome analysis for this complex, nematode-vectored Rathayibacter genera that compares the three agronomically-damaging toxigenic and atoxigenic Rathayibacter species, R. toxicus, R. iranicus, and R. tritici. The exoproteomic comparison identified 1,423 unique proteins between the three species using LC-MS/MS, leading to the identification of putative pathogenicity-related proteins and proteins that may mediate nematode attachment. Of the uniquely identified proteins, 94 homologous proteins were conserved between the three Rathayibacter exoproteomes and comprised between 43.4 – 58.6% of total protein abundance. Comparative analyses revealed both conserved and uniquely expressed extracellular proteins, which, interestingly, had more similarities to extracellular proteins commonly associated with bacterial animal pathogens than classical plant pathogens. This comparative exoproteome analysis will facilitate the characterization of proteins essential for vector attachment and host colonization and assist in the development of serological diagnostic assays.

scholarly journals Scanning Electron-Assisted Dielectric Microscopy Reveals Autophagosome Formation by LC3 and ATG12 in Cultured Mammalian Cells

2021 ◽  
Vol 22 (4) ◽  
pp. 1834
Author(s):  
Tomoko Okada ◽  
Toshihiko Ogura
Keyword(s):  
Mammalian Cells ◽  
Culture Medium ◽  
Related Gene ◽  
Autophagosome Formation ◽  
Intact Cells ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
The Difference ◽  
Related Proteins ◽  
Scanning Electron

Autophagy is an intracellular self-devouring system that plays a central role in cellular recycling. The formation of functional autophagosomes depends on several autophagy-related proteins, including the microtubule-associated proteins 1A/1B light chain 3 (LC3) and the conserved autophagy-related gene 12 (Atg12). We have recently developed a novel scanning electron-assisted dielectric microscope (SE-ADM) for nanoscale observations of intact cells. Here, we used the SE-ADM system to observe LC3- and Atg12-containing autophagosomes in cells labelled in the culture medium with antibodies conjugated to colloidal gold particles. We observed that, during autophagosome formation, Atg12 localized along the actin meshwork structure, whereas LC3 formed arcuate or circular alignments. Our system also showed a difference in the distribution of LC3 and Atg12; Atg12 was broadly distributed while LC3 was more localized. The difference in the spatial distribution demonstrated by our system explains the difference in the size of fluorescent spots due to the fluorescently labelled antibodies observed using optical microscopy. The direct SE-ADM observation of cells should thus be effective in analyses of autophagosome formation.

scholarly journals THE TIME OF SYNTHESIS AND THE CONSERVATION OF MITOSIS-RELATED PROTEINS IN CULTURED HUMAN AMNION CELLS

1967 ◽  
Vol 34 (1) ◽  
pp. 97-110 ◽  
Author(s):  
Jesse E. Sisken ◽  
Elaina Wilkes
Keyword(s):  
Time Lapse ◽  
Major Effect ◽  
Human Amnion ◽  
Daughter Cells ◽  
Low Concentrations ◽  
Associated Proteins ◽  
A Cell ◽  
Quantitative Analyses ◽  
Related Proteins ◽  
Kinetics Of

p-Fluorophenylalanine (PFPA), an analogue of phenylalanine which may be incorporated into proteins, increases the duration of mitosis. In the present experiments, based upon quantitative analyses of time-lapse cinemicrographic films, brief treatments of cells with PFPA are shown to affect the duration of metaphase in only those cells which enter division during or shortly after treatment. The offspring of cells with prolonged metaphases also tend to have prolonged metaphases. Analyses of the kinetics of the appearance of prolonged metaphases indicate that some protein specifically associated with mitosis is synthesized primarily during a period which corresponds closely to G2. The manner in which the defect is passed on to daughter cells indicates that the protein involved is conserved and reutilized by daughter cells for their subsequent divisions. Comparable experiments performed with low concentrations of puromycin indicate that the major effect of PFPA is due to its incorporation into protein rather than its ability to inhibit protein synthesis. The fact that puromycin-induced effects can also be passed on to daughter cells is interpreted to mean that cells make only specific amounts of some mitosis-associated proteins and that if a cell "inherits" a deficiency in such protein it is not able to compensate for the deficiency.

Multi-omics reveals a relationship between endometrial amino acid metabolism and autophagy in women with recurrent miscarriage

2021 ◽  
Author(s):  
Ling Hong ◽  
Yuan chang Zhu ◽  
Su Liu ◽  
Tonghua Wu ◽  
Yuye Li ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Metabolism ◽  
Acid Metabolism ◽  
Recurrent Miscarriage ◽  
Normal Pregnancy ◽  
Beclin 1 ◽  
Endometrial Stromal Cells ◽  
Associated Proteins ◽  
Glutamine Deprivation ◽  
Related Proteins

Abstract Deterioration of the endometrial environment is an essential cause of recurrent miscarriage (RM). However, current studies in terms of endometrial amino acid metabolic characterization and autophagy are still inadequate. This study aimed to assess differences in the endometrial amino acid metabolism and autophagy between normal pregnant and RM women to reveal the association between amino acid metabolism, autophagy, and the endometrial microenvironment. We tried to (1) identify the alternation in metabolite profiles in the RM endometrium; (2) investigate the expression of autophagy-related proteins in RM; (3) elucidate the association between amino acid metabolism and autophagy in RM. Our results showed that glutamine metabolites were upregulated while glutamate metabolites were downregulated in the endometrium of RM women. The levels of autophagy-associated proteins, including LC3B, ATG12, and Beclin-1, were significantly higher in the endometrium of RM women. Hemostasis, autophagy and IFNα signaling were the top three differentially activated signaling pathways between women with RM and normal pregnancy. Interestingly, we also found that the expression of AMPK and GCN2 was significantly up-regulated in the endometrium of women with RM, and the same expression trend was also observed in the human endometrial stromal cells cultured in glutamine deprivation medium. Furthermore, inhibition of AMPK decreased the level of GCN2, indicating a positive correlation between GCN2 and AMPK. The expression of GCN2 was consistent with the expression of ATG12 and beclin-1; however, it was opposite to that of p62. Exposure to glutamine deprivation increased the level of LC3B, GCN2, ATG12 and beclin-1. Altogether, these findings suggested significant crosstalk between amino acid metabolism and autophagy. In summary, our data suggested that aberrant crosstalk between amino acid metabolism and autophagy may contribute to the impaired endometrial microenvironment of RM. Our study may provide new insight into the diagnosis of recurrent miscarriage due to endometrial factors.

A highly evolutionarily conserved mitochondrial protein is structurally related to the protein encoded by the Escherichia coli groEL gene

1988 ◽  
Vol 8 (1) ◽  
pp. 371-380
Author(s):  
T W McMullin ◽  
R L Hallberg
Keyword(s):  
Escherichia Coli ◽  
Tetrahymena Thermophila ◽  
Mitochondrial Protein ◽  
Cross Reactivity ◽  
Macromolecular Complexes ◽  
E Coli ◽  
Groel Gene ◽  
Evolutionarily Conserved ◽  
Associated Proteins ◽  
Related Proteins

We recently reported that a Tetrahymena thermophila 58-kilodalton (kDa) mitochondrial protein (hsp58) was selectively synthesized during heat shock. In this study, we show that hsp58 displayed antigenic similarity with mitochondrially associated proteins from Saccharomyces cerevisiae (64 kDa), Xenopus laevis (60 kDa), Zea mays (62 kDa), and human cells (59 kDa). Furthermore, a 58-kDa protein from Escherichia coli also exhibited antigenic cross-reactivity to an antiserum directed against the T. thermophila mitochondrial protein. The proteins from S. cerevisiae and E. coli antigenically related to hsp58 were studied in detail and found to share several other characteristics with hsp58, including heat inducibility and the property of associating into distinct oligomeric complexes. The T. thermophila, S. cerevisiae, and E. coli macromolecular complexes containing these related proteins had similar sedimentation characteristics and virtually identical morphologies as seen with the electron microscope. The distinctive properties of the E. coli homolog to T. thermophila hsp58 indicate that it is most likely the product of the groEL gene.

HSP27 elevated in mild allergic inflammation protects airway epithelium from H2SO4 effects

1997 ◽  
Vol 273 (2) ◽  
pp. L401-L409 ◽  
Author(s):  
A. T. Hastie ◽  
K. B. Everts ◽  
J. Zangrilli ◽  
J. R. Shaver ◽  
M. B. Pollice ◽  
...  
Keyword(s):  
Airway Epithelium ◽  
Stress Proteins ◽  
Stress Protein ◽  
Allergic Inflammation ◽  
Allergen Challenge ◽  
Bronchial Epithelium ◽  
Ciliary Activity ◽  
Normal Epithelium ◽  
Adverse Conditions ◽  
Before And After

Inflammation in allergic individuals is hypothesized to elevate stress proteins [heat shock proteins (HSP)] in airway epithelium, which may protect cells from further adverse conditions. Allergic, either asthmatic or not, and normal volunteers participated in a 2-day segmental allergen challenge bronchoscopic procedure. Bronchial epithelium was obtained before and after challenge. Epithelium was exposed to medium with H2SO4 (pH5), returned to medium at pH 7.4, and finally harvested for Western blotting with anti-27-kDa HSP (HSP27) antibody. Prechallenge epithelium of all subjects had significantly inhibited ciliary function by H2SO4 (pH 5) conditions (P < 0.001); only epithelium of normals recovered (P = 0.02). Allergic subjects with mild inflammation (< 50 micrograms/ml increase in albumin in bronchoalveolar lavage) had significantly increased HSP27 postchallenge (P = 0.01) and little ciliary dysfunction at pH 5, whereas subjects with severe inflammation (> 50 micrograms/ml increase in albumin) had little change in HSP27 and significant ciliary inhibition (P = 0.02). Normal epithelium had similar trends in HSP27 and equivalent inhibition of ciliary activity at pH 5 before and after allergen challenge. These data indicate that mild inflammation to allergen elevates HSP27 stress protein levels, thereby potentially protecting epithelial function from additional adverse conditions.

scholarly journals The oncornavirus glycoprotein gp69/71: a constituent of the surface of normal and malignant thymocytes.

1975 ◽  
Vol 141 (1) ◽  
pp. 172-187 ◽  
Author(s):  
B C Del Vellano ◽  
B Nave ◽  
B P Croker ◽  
R A Lerner ◽  
F J Dixon
Keyword(s):  
Leukemia Virus ◽  
Molecular Size ◽  
Size Class ◽  
Antigenic Determinants ◽  
Lymphoma Cells ◽  
Differentiated Cells ◽  
Interesting Difference ◽  
Associated Proteins ◽  
Related Proteins ◽  
The Relationship

The oncornavirus related proteins associated with the surface of normal and malignant thymocytes were studied. Three virion-associated proteins (gp69/71, p45, p30) were associated with lymphoma cells from about 70% of the tumors studied. Two virion-associated proteins (gp69/71 and p45 were associated with normal thymocytes form some but not all strains of mice. In gp69/71- mice, conversion to the gp69/71+ phenotype accompanied leukemogenesis. An interesting difference in the apparent molecular size of virus related antigens of the 70,000 dalton size class was detected in lymphoma cells present in involved spleens as compared to involved thymuses. Mice infected as neonates with Scripps leukemia virus make antibody to gp69/71 and some make antibodies to molecules associated with the surface of their own tumors. The significance of the restricted presence of antigens coded for by the viral genome to the surface of some differentiated cells is discussed in reference to (a) the relationship between virion, leukemia associated, and differentiation dependent markers, and (b) the possible consequence to the host of having similar antigenic determinants on three independent structures with replicative potential (virus, normal thymocytes, and tumor cells).

scholarly journals Diversity of the Brain Dystrophin-Glycoprotein Complex

2002 ◽  
Vol 2 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Kevin Culligan ◽  
Kay Ohlendieck
Keyword(s):  
Neuromuscular Disorder ◽  
Glycoprotein Complex ◽  
Associated Proteins ◽  
Dystrophin Glycoprotein Complex ◽  
Dmd Gene ◽  
Related Proteins ◽  
Dystrophin Glycoprotein ◽  
Molecular Brain ◽  
Specific Membrane ◽  
The Brain

Duchenne muscular dystrophy (DMD), the most common inherited neuromuscular disorder, is characterized by progressive muscle wasting and weakness. One third of Duchenne patients suffer a moderate to severe, nonprogressive form of mental retardation. Mutations in the DMD gene are thought to be responsible, with the shorter isoforms of dystrophin implicated in its molecular brain pathogenesis. It is becoming clear that region-specific variations in dystrophin isoforms delegate the composition of the dystrophin-glycoprotein complex in brain, and hence, the function of the specific membrane assembly. Here we summarize the recent advances in the understanding of brain dystrophin, dystrophin-related proteins and dystrophin-associated proteins.

scholarly journals Identification of Cell Wall-Associated Proteins from Phytophthora ramorum

2006 ◽  
Vol 19 (12) ◽  
pp. 1348-1358 ◽  
Author(s):  
Harold J. G. Meijer ◽  
Peter J. I. van de Vondervoort ◽  
Qing Yuan Yin ◽  
Chris G. de Koster ◽  
Frans M. Klis ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Plant Pathogens ◽  
Sodium Dodecyl ◽  
Phytophthora Ramorum ◽  
Extracellular Proteins ◽  
Glycoside Hydrolases ◽  
Secretory Proteins ◽  
Plant Infection ◽  
Associated Proteins

The oomycete genus Phytophthora comprises a large group of fungal-like plant pathogens. Two Phytophthora genomes recently have been sequenced; one of them is the genome of Phytophthora ramorum, the causal agent of sudden oak death. During plant infection, extracellular proteins, either soluble secreted proteins or proteins associated with the cell wall, play important roles in the interaction with host plants. Cell walls of P. ramorum contain 1 to 1.5% proteins, the remainder almost exclusively being accounted for by glucan polymers. Here, we present an inventory of cell-wall-associated proteins based on mass spectrometric sequence analysis of tryptic peptides obtained by proteolytic digestion of sodium dodecyl sulfate-treated mycelial cell walls. In total, 17 proteins were identified, all of which are authentic secretory proteins. Functional classification based on homology searches revealed six putative mucins or mucin-like proteins, five putative glycoside hydrolases, two transglutaminases, one annexin-like protein, the elicitin protein RAM5, one protein of unknown function, and one Kazal-type protease inhibitor. We propose that the cell wall proteins thus identified are important for pathogenicity.

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