Mapping and identification of Brucella melitensis proteins by two-dimensional electrophoresis and microsequencing

✍ Scribed by Ana P. Teixeira-Gomes; Axel Cloeckaert; Guy Bézard; Gérard Dubray; Dr. Michel S. Zygmunt

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 712 KB
• Volume: 18
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150180128

✦ Synopsis

Mapping and identification of Brucella melitensis proteins by two-dimensional electrophoresis and microsequencing

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 1OK-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.