A proteome reference map for <b><i>Vibrio cholerae</i></b> El Tor

Coelho, Ana; Santos, Eidy de Oliveira; Faria, Mauro Luiz da Hora; Carvalho, de; Soares, Márcia Regina; Krüger, Wanda M. A. von; Bisch, Paulo Mascarello

doi:10.1002/pmic.200300685

Cited by 35 publications

(17 citation statements)

References 37 publications

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“…SDS-polyacrylamide gel electrophoresis was performed on a 12% polyacrylamide gel in a Ruby SE600 electrophoresis system (GE Healthcare Biosciences AB). All the bands were excised from three replicate gels as described previously (Hellman et al, 1995;Coelho et al, 2004). Sequencing grade porcine trypsin (Promega, Madison, WI, USA) was used for protein digestion.…”

Section: Secretome Analysesmentioning

confidence: 99%

Genomic and proteomic analyses of the coral pathogen Vibrio coralliilyticus reveal a diverse virulence repertoire

et al. 2011

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Vibrio coralliilyticus has been implicated as an important pathogen of coral species worldwide. In this study, the nearly complete genome of Vibrio coralliilyticus strain P1 (LMG23696) was sequenced and proteases implicated in virulence of the strain were specifically investigated. The genome sequence of P1 (5 513 256 bp in size) consisted of 5222 coding sequences and 58 RNA genes (53 tRNAs and at least 5 rRNAs). Seventeen metalloprotease and effector (vgrG, hlyA and hcp) genes were identified in the genome and expressed proteases were also detected in the secretome of P1. As the VcpA zinc-metalloprotease has been considered an important virulence factor of V. coralliilyticus, a vcpA deletion mutant was constructed to evaluate the effect of this gene in animal pathogenesis. Both wild-type and mutant (DvcpA) strains exhibited similar virulence characteristics that resulted in high mortality in Artemia and Drosophila pathogenicity bioassays and strong photosystem II inactivation of the coral dinoflagellate endosymbiont (Symbiodinium). In contrast, the DvcpA mutant demonstrated higher hemolytic activity and secreted 18 proteins not secreted by the wild type. These proteins included four types of metalloproteases, a chitinase, a hemolysin-related protein RbmC, the Hcp protein and 12 hypothetical proteins. Overall, the results of this study indicate that V. coralliilyticus strain P1 has a diverse virulence repertoire that possibly enables this bacterium to be an efficient animal pathogen.

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Section: Secretome Analysesmentioning

confidence: 99%

Genomic and proteomic analyses of the coral pathogen Vibrio coralliilyticus reveal a diverse virulence repertoire

et al. 2011

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“…2-D gel image database (SWISS-2DPAGE) was established in 1993, with the premise of having an internet-accessible resource where all the spots on high quality 2-D gels of an organism's proteome were clearly annotated and identified (Appel et al, 1993 Regula et al, 2000;Cordwell et al, 2002;Liao et al, 2003;Coelho et al, 2004;Eymann et al, 2004;Folio et al, 2004;Rosen et al, 2004;Encheva et al, 2005;Wang et al, 2005;Encheva et al, 2006). Through concerted and exhaustive efforts, it is often possible to resolve and identify more than a thousand protein spots in a comprehensive 2-D reference map.…”

Section: Whole Cell Proteomementioning

confidence: 99%

Two-dimensional gel electrophoresis in bacterial proteomics

et al. 2012

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Two-dimensional gel electrophoresis (2-DE) is a gel-based technique widely used for analyzing the protein composition of biological samples. It is capable of resolving complex mixtures containing more than a thousand protein components into individual protein spots through the coupling of two orthogonal biophysical separation techniques: isoelectric focusing (first dimension) and polyacrylamide gel electrophoresis (second dimension). 2-DE is ideally suited for analyzing the entire expressed protein complement of a bacterial cell: its proteome. Its relative simplicity and good reproducibility have led to 2-DE being widely used for exploring proteomics within a wide range of environmental and medically-relevant bacteria. Here we give a broad overview of the basic principles and historical development of gel-based proteomics, and how this powerful approach can be applied for studying bacterial biology and physiology. We highlight specific 2-DE applications that can be used to analyze when, where and how much proteins are expressed. The links between proteomics, genomics and mass spectrometry are discussed. We explore how proteomics involving tandem mass spectrometry can be used to analyze (post-translational) protein modifications or to identify proteins of unknown origin by de novo peptide sequencing. The use of proteome fractionation techniques and non-gel-based proteomic approaches are also discussed. We highlight how the analysis of proteins secreted by bacterial cells (secretomes or exoproteomes) can be used to study infection processes or the immune response. This review is aimed at non-specialists who wish to gain a concise, comprehensive and contemporary overview of the nature and applications of bacterial proteomics.

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“…Vc's proteome has been published, but it has not been linked to the human immunome (available Ab specificities), nor has it been tested against a comprehensive panel of Vc convalescent sera or sera of those who are hyperimmune to cholera [42]. While the in vivo expressed OM proteome is a place to start the immunome analysis, the presence of Vc-specific Abs in cholera patients does not prove a role in protection.…”

Section: Stool-associatedmentioning

confidence: 99%