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Mapping and identification of Brucella melitensis proteins by two‐dimensional electrophoresis and microsequencing
Author(s) -
TeixeiraGomes Ana P.,
Cloeckaert Axel,
Bézard Guy,
Dubray Gérard,
Zygmunt Michel S.
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150180128
Subject(s) - groel , microbiology and biotechnology , biochemistry , ribosomal protein , biology , gel electrophoresis , protein subunit , isoelectric point , groes , phosphoprotein , brucella melitensis , chemistry , escherichia coli , brucella , ribosome , enzyme , gene , rna , virology , brucellosis
Two‐dimensional (2‐D) gel electrophoresis was used to map Brucella melitensis proteins. The 2‐D proteins map of B. melitensis B115 revealed 595 silverstained protein spots separated by both isoelectric point and molecular mass. Twenty‐five proteins were identified either by immunoblotting using monoclonal antibodies (MAbs) or by N ‐terminal microsequencing. The protein spots identified by MAbs were the 89 kDa outer membrane protein, DnaK, bacterioferritin, CP24, and BP26. Some spots were identified by N ‐terminal microsequencing as proteins whose sequences had been reported previously from Brucella , such as three heat‐shock proteins, namely DnaK, GroEL and GroES; bacterioferritin; Cu‐Zn superoxide dismutase; and the 50S ribosomal protein L7/L12. Other proteins had amino acid sequences homologous with those of various proteins from other bacteria found in protein databases: ClpP; the 10K‐S protein; the ORFU phosphoprotein; succinyl‐CoA synthetase alpha subunit; an inorganic pyrophosphatase; the Fe and/or Mn superoxide dismutase; the nucleoside diphosphate kinase, an amino acid ABC type transporter, and an electron transfer flavoprotein small subunit. Seven proteins were identified with N ‐terminal sequences not yet reported in databases. The 2‐D map established in this study will be the basis for comparative studies of protein expression in Brucella .