Proteome profile of a pandemic<i>Vibrio parahaemolyticus</i>SC192 strain in the planktonic and biofilm condition

Dharmaprakash, Akhilandeswarre; Mutt, Eshita; Jaleel, Abdul; Sowdhamini, R.; Thomas, Sabu

doi:10.1080/08927014.2014.916696

Cited by 16 publications

(7 citation statements)

References 26 publications

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“…Three biological replicates and three technical replicates were used in this experiment. 100 μg of protein (1 μg/μl) from each sample was made up to 100 μl with 50 mM Ammonium bicarbonate and subjected to disulfide reduction, alkylation and in-solution tryptic digestion as followed by previous publication [44]. The digested peptide solution was centrifuged at maximum rpm at 4 °C for 12 min and the supernatant was stored at −20°C until the analysis using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).…”

Section: Methodsmentioning

confidence: 99%

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Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants

et al. 2019

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Background Enterococcus faecalis is a major clinically relevant nosocomial bacterial pathogen frequently isolated from polymicrobial infections. The biofilm forming ability of E. faecalis attributes a key role in its virulence and drug resistance. Biofilm cells are phenotypically and metabolically different from their planktonic counterparts and many aspects involved in E. faecalis biofilm formation are yet to be elucidated. The strain E. faecalis SK460 used in the present study is esp (Enterococcal surface protein) and fsr (two-component signal transduction system) negative non-gelatinase producing strong biofilm former isolated from a chronic diabetic foot ulcer patient. We executed a label-free quantitative proteomic approach to elucidate the differential protein expression pattern at planktonic and biofilm stages of SK460 to come up with potential determinants associated with Enterococcal biofilm formation. Results The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of proteomic data revealed that biofilm cells expressed higher levels of proteins which are associated with glycolysis, amino acid biosynthesis, biosynthesis of secondary metabolites, microbial metabolism in diverse environments and stress response factors. Besides these basic survival pathways, LuxS-mediated quorum sensing, arginine metabolism, rhamnose biosynthesis, pheromone and adhesion associated proteins were found to be upregulated during the biofilm transit from planktonic stages. The selected subsets were validated by quantitative real-time PCR. In silico functional interaction analysis revealed that the genes involved in upregulated pathways pose a close molecular interaction thereby coordinating the regulatory network to thrive as a biofilm community. Conclusions The present study describes the first report of the quantitative proteome analysis of an esp and fsr negative non gelatinase producing E. faecalis . Proteome analysis evidenced enhanced expression of glycolytic pathways, stress response factors, LuxS quorum signaling system, rhamnopolysaccharide synthesis and pheromone associated proteins in biofilm phenotype. We also pointed out the relevance of LuxS quorum sensing and pheromone associated proteins in the biofilm development of E. faecalis which lacks the Fsr quorum signaling system. These validated biofilm determinants can act as potential inhibiting targets in Enterococcal infections. Electronic supplementary material The online version of this article (10.1186/s12866-019-1527-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

“…All analyses were performed using positive mode ESI using a NanoLockSpray™source. The detailed operational set up was followed as in previous publications [44].…”

Section: Methodsmentioning

confidence: 99%

Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants

et al. 2019

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“…Peptides eluted from the nano-LC were subjected to mass spectrometric analysis on a SYNAPT ® G2 High Definition MS™ System (Waters, UK). The parameters were used as described in Dharmaprakash et al (2014). The mass spectrometer was operated in the “resolution mode” with a resolving power of 18,000 FWHM and the data was obtained in “continuum” format.…”

Section: Methodsmentioning

confidence: 99%

Global Proteomic Response of Caenorhabditis elegans Against PemKSa Toxin

Mir

Balamurugan

2019

Front. Cell. Infect. Microbiol.

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Bacterial exotoxins are major causative agents that infect by promoting cell and tissue damages through disabling the invading host immune system. However, the mode of action by which toxins modulate host immune system and lead cell death is still not completely understood. The nematode, Caenorhabditis elegans has been used as an attractive model host for toxicological studies. In this regard, the present study was undertaken to assess the impact of Staphylococcus aureus toxin (PemK) on the host C. elegans through global proteomics approach. Our proteomic data obtained through LC-MS/MS, subsequent bioinformatics and biochemical analyses revealed that in response to PemK Sa a total of 601 proteins of C. elegans were differentially regulated in response to PemK Sa . The identified proteins were found to mainly participate in ATP generation, protein synthesis, lipid synthesis, cytoskeleton, heat shock proteins, innate immune defense, stress response, neuron degeneration, and muscle assembly. Current findings suggested that involvement of several regulatory proteins that appear to play a role in various molecular functions in combating PemK Sa toxin-mediated microbial pathogenicity and/or host C. elegans immunity modulation. The results provided a preliminary view of the physiological and molecular response of a host toward a toxin and provided insight into highly complex host-toxin interactions.

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“…Biofilm staining of Gram‐positive and Gram‐negative biofilm‐forming isolates with Syto9 (Invitrogen) was carried out as described previously . The slides were then observed under 60× objective using a Nikon Eclipse Ti Confocal Laser scanning inverted microscope (Nikon).…”

Section: Methodsmentioning

confidence: 99%

Metataxonomic approach to decipher the polymicrobial burden in diabetic foot ulcer and its biofilm mode of infection

Suryaletha

John

Radhakrishnan

et al. 2018

International Wound Journal

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Chronic diabetic foot is a global burden affecting millions of people, and the chronicity of an ulcer is directly linked to the diverse bacterial burden and its biofilm mode of infection. The bacterial diversity of 100 chronic diabetic ulcer samples was profiled via traditional culturing method as well as metagenomic approach by sequencing the 16S rRNA V3 hyper-variable region on Illumina Miseq Platform (Illumina, Inc., San Diego, CA). All the relevant clinical metadata, including duration of diabetes, grade of ulcer, presence of neuropathy, and glycaemic level, were noted and correlated with the microbiota. The occurrence and establishment of bacterial biofilm over chronic wound tissues was revealed by Fluorescent in situ Hybridization and Scanning Electron Microscopy. The biofilm-forming ability of predominant bacterial isolates was studied via crystal violet assay and Confocal Laser Scanning Microscopy. The dominant phyla obtained from bacterial diversity analysis were Firmicutes, Proteobacteria, and Actinobacteria. The dominant aerobic pathogens identified by culture method are Pseudomonas, Proteus, Enterococcus, and Staphylococcus, whereas high-throughput sequencing revealed heightened levels of Streptococcus and Corynebacterium along with 22 different obligate anaerobes. The biofilm occurrence in chronic diabetic ulcer infection is well analysed. Herein, we illustrate the comprehensive pattern of bacterial infection and identify the community composition of chronic wound pathogenic biofilm.

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Proteome profile of a pandemicVibrio parahaemolyticusSC192 strain in the planktonic and biofilm condition

Cited by 16 publications

References 26 publications

Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants

Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants

Global Proteomic Response of Caenorhabditis elegans Against PemKSa Toxin

Metataxonomic approach to decipher the polymicrobial burden in diabetic foot ulcer and its biofilm mode of infection

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