scholarly journals Search of Potential Vaccine Candidates against Trueperella pyogenes Infections through Proteomic and Bioinformatic Analysis

Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 314 ◽  
Author(s):  
Ángela Galán-Relaño ◽  
Lidia Gómez-Gascón ◽  
Antonio Rodríguez-Franco ◽  
Inmaculada Luque ◽  
Belén Huerta ◽  
...  
Keyword(s):  
Cell Wall ◽  
Membrane Proteins ◽  
Bioinformatic Analysis ◽  
Surface Proteins ◽  
Live Cells ◽  
Economic Losses ◽  
Diagnostic Tools ◽  
Cell Wall Proteins ◽  
Vaccine Candidates ◽  
Trueperella Pyogenes

Trueperella pyogenes is an opportunistic pathogen, responsible for important infections in pigs and significant economic losses in swine production. To date, there are no available commercial vaccines to control diseases caused by this bacterium. In this work, we performed a comparative proteomic analysis of 15 T. pyogenes clinical isolates, by “shaving” live cells, followed by LC-MS/MS, aiming at the identification of the whole set of surface proteins (i.e., the “pan-surfome”) as a source of antigens to be tested in further studies as putative vaccine candidates, or used in diagnostic tools. A total of 140 surface proteins were detected, comprising 25 cell wall proteins, 10 secreted proteins, 23 lipoproteins and 82 membrane proteins. After describing the “pan-surfome”, the identified proteins were ranked in three different groups based on the following criteria: to be (i) surface-exposed, (ii) highly conserved and (iii) widely distributed among different isolates. Two cell wall proteins, three lipoproteins, four secreted and seven membrane proteins were identified in more than 70% of the studied strains, were highly expressed and highly conserved. These proteins are potential candidates, alone or in combination, to obtain effective vaccines against T. pyogenes or to be used in the diagnosis of this pathogen.

scholarly journals Proteomics-Based Identification of Anchorless Cell Wall Proteins as Vaccine Candidates against Staphylococcus aureus

2009 ◽  
Vol 77 (7) ◽  
pp. 2719-2729 ◽  
Author(s):  
Eva Glowalla ◽  
Bettina Tosetti ◽  
Martin Krönke ◽  
Oleg Krut
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Survival Rates ◽  
Proteome Analysis ◽  
Surface Proteins ◽  
Human Neutrophils ◽  
Cell Wall Proteins ◽  
Lethal Challenge ◽  
Vaccine Candidates

ABSTRACT Staphylococcus aureus is an important human pathogen with increasing clinical impact due to the extensive spread of antibiotic-resistant strains. Therefore, development of a protective polyvalent vaccine is of great clinical interest. We employed an intravenous immunoglobulin (IVIG) preparation as a source of antibodies directed against anchorless S. aureus surface proteins for identification of novel vaccine candidates. In order to identify such proteins, subtractive proteome analysis (SUPRA) of S. aureus anchorless cell wall proteins was performed. Proteins reacting with IVIG but not with IVIG depleted of S. aureus-specific opsonizing antibodies were considered vaccine candidates. Nearly 40 proteins were identified by this preselection method using matrix-assisted laser desorption ionization—time of flight analysis. Three of these candidate proteins, enolase (Eno), oxoacyl reductase (Oxo), and hypothetical protein hp2160, were expressed as glutathione S-transferase fusion proteins, purified, and used for enrichment of corresponding immunoglobulin Gs from IVIG by affinity chromatography. Use of affinity-purified anti-Eno, anti-Oxo, and anti-hp2160 antibodies resulted in opsonization, phagocytosis, and killing of S. aureus by human neutrophils. High specific antibody titers were detected in mice immunized with recombinant antigens. In mice challenged with bioluminescent S. aureus, reduced staphylococcal spread was measured by in vivo imaging. The recovery of S. aureus CFU from organs of immunized mice was diminished 10- to 100-fold. Finally, mice immunized with hp2160 displayed statistically significant higher survival rates after lethal challenge with clinically relevant S. aureus strains. Taken together, our data suggest that anchorless cell wall proteins might be promising vaccine candidates and that SUPRA is a valuable tool for their identification.

scholarly journals Flagellar adhesion in chlamydomonas induces synthesis of two high molecular weight cell surface proteins

1983 ◽  
Vol 96 (3) ◽  
pp. 589-597 ◽  
Author(s):  
WJ Snell ◽  
A Clausell ◽  
WS Moore
Keyword(s):  
Cell Wall ◽  
Cell Surface ◽  
Adhesion Molecules ◽  
Mating Type ◽  
Surface Proteins ◽  
Cell Wall Proteins ◽  
Intact Cells ◽  
Mating Reaction ◽  
Ionic Detergent ◽  
Flagellar Proteins

Because our previous studies (Snell, W.J., and W.S. Moore, 1980, J. Cell Biol. 84:203- 210) on the mating reaction of chlamydomonas reinhardtii showed that there was an adhesion-induced turnover of proteins whose synthesis is induced during aggregation. Analysis by SDS PAGE and autoradiography showed that proteins of 220,000 M(r) and 165, 000 M(r) (designated A(1) and A(2) respectively) consistently showed a high rate of synthesis only in flagella or flagellar membrane-enriched fractions prepared from aggregating gametes. Since the two proteins were soluble in the non-ionic detergent NP-40 and were removed from intact cells by a brief pronase treatment, it is likely that A(1) and A(2) are membrane proteins expose on the cell surface. A(1) and A(2) were each synthesized by gametes of both mating types (mt(-) and mt(+)) and synthesis of these two proteins could be detected in the normal mating reaction (wild type mt(-) and mt(+)), in mixtures of mt(-) and impotent mt(+) gametes (which could aggregate but not fuse), and in mixtures of gametes of a single mating type with isolated flagella of the opposite mating type. Cells aggregating in tunicamycin, an inhibitor of protein glycosylation, lost their adhesiveness during aggregation and did not synthesize the 220,000 M(r) protein but instead produced a protein (possibly an underglycosylated form of A(1)) of slightly lower mol wt. The 220,000 and 165,000 M(R) proteins appeared to be flagellar proteins and not cell wall proteins because A(1) and A(2) did not co-migrate with previously identified cell wall proteins, and synthesis of the two proteins could not be detected in flagella-less (bald-2) mutant cells. Analysis of the adhesive activity of sucrose gradient fraction of detergent (octyl glucoside)-solubilized flagellar membranes revealed that fractions containing A(1) and A(2) did not have detectable adhesive activity. The possibility remains that A(1) and A(2) are adhesion molecules whose activity could not be measured in the assay we used. Alternatively, the 220,000 and 165,000 M(r) proteins may be inactivated adhesion molecules or else they may be flagellar surface proteins involved only indirectly in the adhesion process.

scholarly journals Proteomic and Bioinformatic Analysis of Streptococcus suis Human Isolates: Combined Prediction of Potential Vaccine Candidates

Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 188 ◽  
Author(s):  
Esther Prados de la Torre ◽  
Antonio Rodríguez-Franco ◽  
Manuel J. Rodríguez-Ortega
Keyword(s):  
Streptococcus Suis ◽  
Bioinformatic Analysis ◽  
Surface Proteins ◽  
Clinical Isolates ◽  
Economic Losses ◽  
Bacterial Cells ◽  
Label Free ◽  
Bioinformatic Tools ◽  
Potential Vaccine ◽  
Live Bacterial Cells

Streptococcus suis is a Gram-positive bacterium responsible for major infections in pigs and economic losses in the livestock industry, but also an emerging zoonotic pathogen causing serious diseases in humans. No vaccine is available so far against this microorganism. Conserved surface proteins are among the most promising candidates for new and effective vaccines. Until now, research on this pathogen has focused on swine isolates, but there is a lack of studies to identify and characterize surface proteins from human clinical isolates. In this work, we performed a comparative proteomic analysis of six clinical isolates from human patients, all belonging to the major serotype 2, by “shaving” the live bacterial cells with trypsin, followed by LC-MS/MS analysis. We identified 131 predicted surface proteins and carried out a label-free semi-quantitative analysis of protein abundances within the six strains. Then, we combined our proteomics results with bioinformatic tools to help improving the selection of novel antigens that can enter the pipeline of vaccine candidate testing. Our work is then a complement to the reverse vaccinology concept.

scholarly journals Comparison of Polyclonal Antibodies to Three Different Preparations of Mycobacterium Paratuberculosis in Immunohistochemical Diagnosis of Johne's Disease in Cattle

1996 ◽  
Vol 8 (4) ◽  
pp. 469-473 ◽  
Author(s):  
Judith R. Stabel ◽  
Mark R. Ackermann ◽  
Jesse P. Goff
Keyword(s):  
Cell Wall ◽  
Polyclonal Antibodies ◽  
Keyhole Limpet Hemocyanin ◽  
Giant Cells ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Diagnostic Tools ◽  
Cell Wall Proteins ◽  
Incomplete Freund’S Adjuvant ◽  
Immunohistochemical Diagnosis

Polyclonal antisera were raised in rabbits against preparations of live and heat-killed Mycobac- terium paratuberculosis and cell-wall proteins of M. paratuberculosis and were evaluated as diagnostic tools in immunohistochemical staining of bovine tissue. Live preparations of M. paratuberculosis (LMp) were inoculated intraperitoneally or intravenously at 109/ml. Heat-killed M. paratuberculosis (HKMp) was prepared by treatment of bacteria at 85 C for 10 minutes. Cell-wall proteins were isolated from M. paratuberculosis and conjugated to keyhole limpet hemocyanin to improve antigenicity (KLH-CWPMp). The HKMp and KLH-CWPMp preparations were emulsified in incomplete Freund's adjuvant before subcutaneous inoculation of rabbits. Antibody titers in the terminal blood sample were higher for HKMp and KLH-CWPMp than for LMp rabbits (1:1,024 vs. 1:64). The KLH-CWPMp antibody did not cross-react with M. bovis-infected tissues. Sensitivity and specificity of immunohistochemical detection of Johne's disease (paratuberculosis) from bovine tissues was much higher for the KLH-CWPMp polyclonal antibody. Immunoreactivity of the antibody resulted in staining of bacteria in the cytoplasm of macrophages, mononuclear giant cells, and extracellular bacteria in both intestine and lymph node.

Do Proline-rich Proteins Modulate a Transglutaminase Catalyzed Mechanism of Candidal Adhesion?

1993 ◽  
Vol 4 (3) ◽  
pp. 293-299 ◽  
Author(s):  
S.D. Bradway ◽  
M.J. Levine
Keyword(s):  
Molecular Weight ◽  
Cell Wall ◽  
High Molecular Weight ◽  
Surface Proteins ◽  
Cross Linking ◽  
Cell Wall Proteins ◽  
Epithelial Surface ◽  
High Molecular Weight Complex ◽  
Sds Page ◽  
Cross Links

Previously, we reported that a membrane-bound epithelial enzyme, transglutaminase (TGase), catalyzes the covalent cross-linking of acidic proline-rich proteins (APRPs) to surface proteins of buccal epithelial cells (BECs). The purpose of this study was twofold: (1) to provide evidence that TGase stabilizes C. albicans adhesion by covalently cross-linking C. albicans and BEC surface proteins and (2) to implicate PRPs in the modulation of this adhesive mechanism. The reactivity of candidal cell wall proteins with TGase was assessed in two separate experiments. Initially, following incubation with native BECs, the cross-linking of iodinated candidal cell wall proteins into high-molecular-weight complexes, as shown by SDS-PAGE/ autoradiography, was inhibited by the TGase inhibitor iodoacetamide. Additionally, [14C]putrescine in the presence of purified TGase, but not [14C]putrescine alone, was shown by SDS-PAGE/fluorography to be cross-linked into surface proteins of both morphogenetic forms (blastospore > hyphal forms) of C. albicans. In adherence assays, a component of both blastospore and hyphal form Candida/BEC adherence was shown to be resistant to detachment by heating adherent cells in 1% SDS at 100°C. However, pretreatment of BECs with iodoacetamide decreased SDS resistant adherence of both forms of C. albicans by =75%. When incubated with [125I]APRPs and purified TGase, both morphogenetic forms of C. albicans bound dramatically more APRP than controls without TGase. [125I]APRP binding in experimental, but not control, samples was resistant to repeated extraction (48 h) with 4% SDS/10% β-mercaptoethanol at 65°C, suggesting that [125I]APRPs were cross-linked to the Candida surface. SDS-PAGE/fluorography was used to verify that APRPs, in Lyticase digests of Candida cell walls, were cross-linked into a high-molecular-weight complex. These experiments suggest that epithelial TGase may stabilize Candida adherence by cross-linking Candida and BEC surface proteins. Additionally, because TGase cross-links APRPs to candidal and epithelial surface proteins, APRPs may interfere with TGase catalyzed mechanisms of adhesion. Supported by USPHS grants DE00185, DE07585, and OSU Seed grant.

scholarly journals Clade-specific chromosomal rearrangements and loss of subtelomeric adhesins in Candida auris

Genetics ◽  
2021 ◽  
Author(s):  
José F Muñoz ◽  
Rory M Welsh ◽  
Terrance Shea ◽  
Dhwani Batra ◽  
Lalitha Gade ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Surface ◽  
Genome Instability ◽  
Chromosomal Rearrangements ◽  
Multidrug Resistant ◽  
Antifungal Drugs ◽  
Surface Proteins ◽  
Cell Wall Proteins ◽  
Candida Auris ◽  
Cell Surface Proteins

Abstract Candida auris is an emerging fungal pathogen of rising concern due to global spread, the ability to cause healthcare-associated outbreaks, and antifungal resistance. Genomic analyses revealed that early contemporaneously detected cases of C. auris were geographically stratified into four major clades. While Clades I, III, and IV are responsible for ongoing outbreaks of invasive and multidrug-resistant infections, Clade II, also termed the East Asian clade, consists primarily of cases of ear infection, is often susceptible to all antifungal drugs, and has not been associated with outbreaks. Here, we generate chromosome-level assemblies of twelve isolates representing the phylogenetic breadth of these four clades and the only isolate described to date from Clade V. This Clade V genome is highly syntenic with those of Clades I, III, and IV, although the sequence is highly divergent from the other clades. Clade II genomes appear highly rearranged, with translocations occurring near GC-poor regions, and large subtelomeric deletions in most chromosomes, resulting in a substantially different karyotype. Rearrangements and deletion lengths vary across Clade II isolates, including two from a single patient, supporting ongoing genome instability. Deleted subtelomeric regions are enriched in Hyr/Iff-like cell-surface proteins, novel candidate cell wall proteins, and an ALS-like adhesin. Cell wall proteins from these families and other drug-related genes show clade-specific signatures of selection in Clades I, III, and IV. Subtelomeric dynamics and the conservation of cell surface proteins in the clades responsible for global outbreaks causing invasive infections suggest an explanation for the different phenotypes observed between clades.

Development of a method for the extraction and analysis of grape skin proteins strongly bound to cell walls

OENO One ◽  
2013 ◽  
Vol 47 (2) ◽  
pp. 129
Author(s):  
Grégory Pasquier ◽  
Delphine Lapaillerie ◽  
Jean-William Dupuy ◽  
Anne-Marie Lomenech ◽  
Stéphane Claverol ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Protein Composition ◽  
Bioinformatic Analysis ◽  
Soluble Proteins ◽  
Cell Wall Proteins ◽  
Grape Skin ◽  
Skin Cell ◽  
Grape Skins ◽  
Cellular Aggregates

<p style="text-align: justify;"><strong>Aim</strong>: To better understand the protein composition of grape skin cell walls, we have developed a method to analyse the strongly bound cell wall proteins.</p><p style="text-align: justify;"><strong>Methods and results</strong>: The protocol was developed with grape skins at full maturity. The critical steps of this protocol were : (i) the elimination of cellular aggregates, (ii) the elimination of soluble proteins, and (iii) the localization of the identified proteins within the cell wall. To verify whether these three conditions were met, the decrease in the quantity of cellular aggregates was followed by optical microscopy, the removal of soluble proteins was measured by chemical assay, and the presence of proteins located in cell walls was demonstrated by extensive bioinformatic analysis. The process made it possible to obtain a four-fold reduction in the amount of cellular aggregates, a reduction in the concentration of soluble proteins below the method detection limit, and a high proportion of proteins predicted to be secreted (79 %).</p><p style="text-align: justify;"><strong>Conclusion</strong>: The protocol described in this paper constitutes the first method to analyse proteins strongly bound to cell walls in grape skins. However, this method excludes the identification of labile proteins or proteins weakly bound to the cell wall.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: This protocol can be used for studying the role that strongly bound cell wall proteins play in development and defense processes in grape skins.</p>

scholarly journals Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii

2010 ◽  
Vol 9 (10) ◽  
pp. 1622-1634 ◽  
Author(s):  
Angela H. Nobbs ◽  
M. Margaret Vickerman ◽  
Howard F. Jenkinson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Wall ◽  
Candida Albicans ◽  
Surface Proteins ◽  
Collagen Type Iv ◽  
Cell Wall Proteins ◽  
Streptococcus Gordonii ◽  
Content Type ◽  
Salivary Pellicle ◽  
Polymicrobial Communities

ABSTRACT Colonization and infection of the human host by opportunistic pathogen Candida albicans derive from an ability of this fungus to colonize mucosal tissues and prosthetic devices within the polymicrobial communities present. To determine the functions of C. albicans cell wall proteins in interactions with host or bacterial molecules, Saccharomyces cerevisiae was utilized as a surrogate host to express C. albicans cell wall proteins Als3p, Eap1p, Hwp1p, and Rbt1p. Salivary pellicle and fibrinogen were identified as novel substrata for Als3p and Hwp1p, while only Als3p mediated adherence of S. cerevisiae to basement membrane collagen type IV. Parental S. cerevisiae cells failed to form biofilms on salivary pellicle, polystyrene, or silicone, but cells expressing Als3p or Hwp1p exhibited significant attachment to each surface. Virulence factor Rbt1p also conferred lower-level binding to salivary pellicle and polystyrene. S. cerevisiae cells expressing Eap1p formed robust biofilms upon polystyrene surfaces but not salivary pellicle. Proteins Als3p and Eap1p, and to a lesser degree Hwp1p, conferred upon S. cerevisiae the ability to bind cells of the oral primary colonizing bacterium Streptococcus gordonii. These interactions, which occurred independently of amyloid aggregate formation, provide the first examples of specific C. albicans surface proteins serving as receptors for bacterial adhesins. Streptococcus gordonii did not bind parental S. cerevisiae or cells expressing Rbt1p. Taken collectively, these data suggest that a network of cell wall proteins comprising Als3p, Hwp1p, and Eap1p, with complementary adhesive functions, promotes interactions of C. albicans with host and bacterial molecules, thus leading to effective colonization within polymicrobial communities.

scholarly journals PREDICTION OF CELL WALL SORTING SIGNALS IN GRAM-POSITIVE BACTERIA WITH A HIDDEN MARKOV MODEL: APPLICATION TO COMPLETE GENOMES

2008 ◽  
Vol 06 (02) ◽  
pp. 387-401 ◽  
Author(s):  
ZOI I. LITOU ◽  
PANTELIS G. BAGOS ◽  
KONSTANTINOS D. TSIRIGOS ◽  
THEODORE D. LIAKOPOULOS ◽  
STAVROS J. HAMODRAKAS
Keyword(s):  
Cell Wall ◽  
Markov Model ◽  
Hidden Markov Model ◽  
Hidden Markov ◽  
Surface Proteins ◽  
Data Sets ◽  
Cell Wall Proteins ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Sorting Signals

Surface proteins in Gram-positive bacteria are frequently implicated in virulence. We have focused on a group of extracellular cell wall-attached proteins (CWPs), containing an LPXTG motif for cleavage and covalent coupling to peptidoglycan by sortase enzymes. A hidden Markov model (HMM) approach for predicting the LPXTG-anchored cell wall proteins of Gram-positive bacteria was developed and compared against existing methods. The HMM model is parsimonious in terms of the number of freely estimated parameters, and it has proved to be very sensitive and specific in a training set of 55 experimentally verified LPXTG-anchored cell wall proteins as well as in reliable data sets of globular and transmembrane proteins. In order to identify such proteins in Gram-positive bacteria, a comprehensive analysis of 94 completely sequenced genomes has been performed. We identified, in total, 860 LPXTG-anchored cell wall proteins, a number that is significantly higher compared to those obtained by other available methods. Of these proteins, 237 are hypothetical proteins according to the annotation of SwissProt, and 88 had no homologs in the SwissProt database — this might be evidence that they are members of newly identified families of CWPs. The prediction tool, the database with the proteins identified in the genomes, and supplementary material are available online at .

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