Analysis of Extracellular Proteome of Staphylococcus aureus: A Mass Spectrometry based Proteomics Method of Exotoxin Characterisation

2020 ◽  
Vol 17 (1) ◽  
pp. 3-9
Author(s):  
Rajdeep Das ◽  
Nisha D`souza ◽  
Surya K. Choubey ◽  
Sethumadhavan Murlidharan ◽  
Anura V. Kurpad ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Extracellular Proteins ◽  
Host Cells ◽  
Dependent Manner ◽  
Extracellular Proteome ◽  
Cytoplasmic Proteins ◽  
Wide Range

Background: Staphylococcus aureus (S. aureus), an important pathogen, causes a wide range of infections in human starting from food poisoning to septicemia. It affects the host cells with various exotoxins, known as virulence factors, which are synthesized in growth phase-dependent manner of the bacteria. S. aureus has been reported to become resistant to antibiotics rapidly. Among two common clinical isolates, Methicillin-sensitive S. aureus (MSSA) and Methicillin-resistant S. aureus (MRSA), MRSA pose major problems across hospitals around the world. Objective: The objective of the present study was to profile the exoproteins of Methicillin-sensitive S. aureus (ATCC 25293) and subsequently to establish a proteomics-based method of characterization of S. aureus that is crucial in treating hospital-acquired infections. Methods: We used two-dimensional nanoLC/ESI-MS based proteomic platform to characterize and quantify the exoproteins isolated from Methicillin-sensitive S. aureus (ATCC 25293) strain. Results: A total of 69 proteins were identified from extracellular proteome pool of ATCC 25293 strain that includes 18 extracellular proteins, 40 cytoplasmic proteins, 2 membrane proteins, 3 cell wall proteins and 6 uncharacterized proteins. Conclusion: We propose that this mass spectrometry-based proteomics method of characterization of exoproteins might be useful to identify S. aureus strains that are resistant to antibiotics.

scholarly journals Cytoplasmic Control of Premature Activation of a Secreted Protease Zymogen: Deletion of Staphostatin B (SspC) in Staphylococcus aureus 8325-4 Yields a Profound Pleiotropic Phenotype

2005 ◽  
Vol 187 (5) ◽  
pp. 1751-1762 ◽  
Author(s):  
Lindsey N. Shaw ◽  
Ewa Golonka ◽  
Grzegorz Szmyd ◽  
Simon J. Foster ◽  
James Travis ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Envelope ◽  
Extracellular Proteins ◽  
Growth Media ◽  
Cytoplasmic Proteins ◽  
Wide Range ◽  
Sigma B ◽  
Intracellular Proteins ◽  
Autolytic Activity ◽  
Total Lack

ABSTRACT The cytoplasmic protein SspC of Staphylococcus aureus, referred to as staphostatin B, is a very specific, tightly binding inhibitor of the secreted protease staphopain B (SspB). SspC is hypothesized to protect intracellular proteins against proteolytic damage by prematurely folded and activated staphopain B (M. Rzychon, A. Sabat, K. Kosowska, J. Potempa, and A. Dubin, Mol. Microbiol. 49:1051-1066, 2003). Here we provide evidence that elimination of intracellular staphopain B activity is indeed the function of SspC. An isogenic sspC mutant of S. aureus 8325-4 exhibits a wide range of striking pleiotropic alterations in phenotype, which distinguish it from the parent. These changes include a defect in growth, a less structured peptidoglycan layer within the cell envelope, severely decreased autolytic activity, resistance to lysis by S. aureus phages, extensively diminished sensitivity to lysis by lysostaphin, the ability to form a biofilm, and a total lack of extracellular proteins secreted into the growth media. The same phenotype was also engineered by introduction of sspB into an 8325-4 sspBC mutant. In contrast, sspC inactivation in the SH1000 strain did not yield any significant changes in the mutant phenotype, apparently due to strongly reduced expression of sspB in the sigma B-positive background. The exact pathway by which these diverse aberrations are exerted in 8325-4 is unknown, but it is apparent that a very small amount of staphopain B (less than 20 ng per 200 μg of cell proteins) is sufficient to bring about these widespread changes. It is proposed that the effects observed are modulated through the proteolytic degradation of several cytoplasmic proteins within cells lacking the inhibitor. Seemingly, some of these proteins may play a role in protein secretion; hence, their proteolytic inactivation by SspB has pleiotropic effects on the SspC-deficient mutant.

Phylogeny Characterization of Seb Gene Encoding Enterotoxins in Staphylococcus aureus Isolated from Raw Milk and Cheese

2019 ◽  
Vol 9 (o3) ◽  
Author(s):  
Fatima N. Aziz ◽  
Laith Abdul Hassan Mohammed-Jawad
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Raw Milk ◽  
Staphylococcal Enterotoxin B ◽  
Human Pathogen ◽  
Conventional Pcr ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Gene Encoding

Food poisoning due to the bacteria is a big global problem in economically and human's health. This problem refers to an illness which is due to infection or the toxin exists in nature and the food that use. Milk is considered a nutritious food because it contains proteins and vitamins. The aim of this study is to detect and phylogeny characterization of staphylococcal enterotoxin B gene (Seb). A total of 200 milk and cheese samples were screened. One hundred ten isolates of Staphylococcus aureus pre-confirmed using selective and differential media with biochemical tests. Genomic DNA was extracted from the isolates and the SEB gene detects using conventional PCR with specific primers. Three staphylococcus aureus isolates were found to be positive for Seb gene using PCR and confirmed by sequencing. Sequence homology showed variety range of identity starting from (100% to 38%). Phylogenetic tree analyses show that samples (6 and 5) are correlated with S. epidermidis. This study discovered that isolates (A6-RLQ and A5-RLQ) are significantly clustered in a group with non- human pathogen Staphylococcus agnetis.

scholarly journals Complement Inactivation Strategy of Staphylococcus aureus Using Decay-Accelerating Factor and the Response of Infected HaCaT Cells

2021 ◽  
Vol 22 (8) ◽  
pp. 4015
Author(s):  
Kyoung Ok Jang ◽  
Youn Woo Lee ◽  
Hangeun Kim ◽  
Dae Kyun Chung
Keyword(s):  
Staphylococcus Aureus ◽  
Immune Cells ◽  
Respiratory Infections ◽  
Food Poisoning ◽  
Surface Expression ◽  
Host Cells ◽  
Hacat Cells ◽  
Decay Accelerating Factor ◽  
Ligand Expression ◽  
The Complement System

Staphylococcus aureus is a species of Gram-positive staphylococcus. It can cause sinusitis, respiratory infections, skin infections, and food poisoning. Recently, it was discovered that S. aureus infects epithelial cells, but the interaction between S. aureus and the host is not well known. In this study, we confirmed S. aureus to be internalized by HaCaT cells using the ESAT-6-like protein EsxB and amplified within the host over time by escaping host immunity. S. aureus increases the expression of decay-accelerating factor (CD55) on the surfaces of host cells, which inhibits the activation of the complement system. This mechanism makes it possible for S. aureus to survive in host cells. S. aureus, sufficiently amplified within the host, is released through the initiation of cell death. On the other hand, the infected host cells increase their surface expression of UL16 binding protein 1 to inform immune cells that they are infected and try to be eliminated. These host defense systems seem to involve the alteration of tight junctions and the induction of ligand expression to activate immune cells. Taken together, our study elucidates a novel aspect of the mechanisms of infection and immune system evasion for S. aureus.

scholarly journals Functional Characterization of a Na+-Coupled Dicarboxylate Carrier Protein from Staphylococcus aureus

2005 ◽  
Vol 187 (15) ◽  
pp. 5189-5194 ◽  
Author(s):  
Jason A. Hall ◽  
Ana M. Pajor
Keyword(s):  
Staphylococcus Aureus ◽  
Structural Analysis ◽  
Transport Properties ◽  
Sequence Homology ◽  
Cytoplasmic Membrane ◽  
Functional Characterization ◽  
Carrier Protein ◽  
Dependent Manner ◽  
Reaction Sequence

ABSTRACT We have cloned and functionally characterized a Na+-coupled dicarboxylate transporter, SdcS, from Staphylococcus aureus. This carrier protein is a member of the divalent anion/Na+ symporter (DASS) family and shares significant sequence homology with the mammalian Na+/dicarboxylate cotransporters NaDC-1 and NaDC-3. Analysis of SdcS function indicates transport properties consistent with those of its eukaryotic counterparts. Thus, SdcS facilitates the transport of the dicarboxylates fumarate, malate, and succinate across the cytoplasmic membrane in a Na+-dependent manner. Furthermore, kinetic work predicts an ordered reaction sequence with Na+ (K 0.5 of 2.7 mM) binding before dicarboxylate (Km of 4.5 μM). Because this transporter and its mammalian homologs are functionally similar, we suggest that SdcS may serve as a useful model for DASS family structural analysis.

scholarly journals Characterization of Staphylococcus aureus strains and evidence for the involvement of non-classical enterotoxin genes in food poisoning outbreaks

2018 ◽  
Vol 365 (13) ◽  
Author(s):  
Laurentiu-Mihai Ciupescu ◽  
Frederic Auvray ◽  
Isabela Madalina Nicorescu ◽  
Thomas Meheut ◽  
Veronica Ciupescu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Enterotoxin Genes

scholarly journals Characteristics of Extracellular Vesicles Released by the Pathogenic Yeast-Like Fungi Candida glabrata, Candida parapsilosis and Candida tropicalis

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1722 ◽  
Author(s):  
Justyna Karkowska-Kuleta ◽  
Kamila Kulig ◽  
Elzbieta Karnas ◽  
Ewa Zuba-Surma ◽  
Olga Woznicka ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Phosphoglycerate Kinase ◽  
Host Cells ◽  
Opportunistic Pathogens ◽  
Candida Spp ◽  
Fungal Cell ◽  
Pathogenic Yeast ◽  
Cytoplasmic Proteins ◽  
Candidal Species

Candida spp. yeast-like fungi are opportunistic pathogens in humans and have been recently found to release extracellular vesicles (EVs) that are involved in many vital biological processes in fungal cells. These include communication between microorganisms and host–pathogen interactions during infection. The production of EVs and their content have been significantly characterized in the most common candidal species Candida albicans, including the identification of numerous virulence factors and cytoplasmic proteins in the EV cargo. We have here conducted the isolation and proteomic characterization of EVs produced by the clinically important non-albicans Candida species C. glabrata, C. tropicalis and C. parapsilosis. With the use of ultracentrifugation of the cell-free culture supernatant, the candidal EVs were collected and found to be a heterogeneous population of particles for each species with sizes ranging from 60–280 nm. The proteinaceous contents of these vesicles were analyzed using LC-MS/MS, with particular attention paid to surface-expressed proteins that would come into immediate and direct contact with host cells. We thereby identified 42 extracellular and surface-connected proteins from C. glabrata, 33 from C. parapsilosis, and 34 from C. tropicalis, including membrane-associated transporters, glycoproteins and enzymes involved in the organization of the fungal cell wall, as well as several cytoplasmic proteins, including alcohol dehydrogenase, enolase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and pyruvate kinase, for which the vesicular transport is a possible mechanism underlying their non-classical secretion.

scholarly journals Deletion of SarX decreases biofilm formation of Staphylococcus aureus in a polysaccharide intercellular adhesin (PIA) dependent manner

2020 ◽  
Author(s):  
Zhihao Hao ◽  
Yinjuan Guo ◽  
Lulin Rao ◽  
Jingyi Yu ◽  
Qing Zhan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Mutant Strain ◽  
Biofilm Formation ◽  
Transcriptional Activity ◽  
Extracellular Proteins ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Biofilm Production ◽  
Fold Reduction ◽  
Operon Expression

Abstract Background: Biofilm formation by Staphylococcus aureus is an important virulence determinant mediated by the polysaccharide intercellular adhere (PIA) encoded by ica operon or by surface and extracellular proteins. Previous studies have shown that S. epidermidis SarX protein regulated the transcriptional activity of the agr and ica loci and controled the biofilm phenotype, primarily by regulating icaADBC transcription and PIA production.Results: In this study, our results indicated that biofilm formation and detachment of S. aureus were significantly decreased in the sarX mutant strain. sarX mutant in S. aureus biofilm formation was related to the production of PIA and not to that of eDNA. RT-PCR results showed that deletion of sarX was associated with a 1.8-fold reduction in spa transcription, which was complemented by sarX. Expression of Spa protein was decreased in sarX mutant strain.Conclusions: sarX promoted biofilm production of S. aureus may mediated primarily through increasing ica operon expression and PIA production. Deletion of sarX was associated with reduction in spa transcription.

scholarly journals Immune System Evasion Mechanisms in Staphylococcus aureus: Current Understanding

2020 ◽  
Vol 14 (4) ◽  
pp. 2219-2234
Author(s):  
Hesham A. Malak ◽  
Hussein H. Abulreesh ◽  
Sameer R. Organji ◽  
Khaled Elbanna ◽  
Mohammed R. Shaaban ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Immune System ◽  
Soft Tissue ◽  
Immune Cells ◽  
Bloodstream Infections ◽  
Host Cells ◽  
Human Pathogen ◽  
Immune Systems ◽  
Wide Range ◽  
New Strategy

Staphylococcus aureus is a major human pathogen that may cause a wide range of infections and is a frequent cause of soft tissue and bloodstream infections. It is a successful pathogen due to its collective virulence factors and its ability to evade the host immune systems. The review aims to highlight how S. aureus destroys and damage the host cells and explains how immune cells can respond to this pathogen. This review may also provide new insights that may be useful for developing new strategy for combating MRSA and its emerging clones such as community-associated methicillin-resistant S. aureus (CA-MRSA).

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