Analysis of protein expression in Brucella abortus mutants with different growth rates by two-dimensional gel electrophoresis and LC-MS/MS peptide analysis

• Published in: Journal of veterinary science (Suwŏn-si, Korea) Vol. 19; no. 2; pp. 216 - 231
• Main Authors: Park, Woo Bin, Im, Young Bin, Shim, Soojin, Yoo, Han Sang
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society of Veterinary Science 01.03.2018; 대한수의학회
• Subjects: Bacterial Proteins - biosynthesis; Brucella abortus - genetics; Brucella abortus - growth & development; Brucella abortus - metabolism; Chromatography, Liquid; Electrophoresis, Gel, Two-Dimensional; Mutagenesis, Insertional; Original; Tandem Mass Spectrometry; 수의학; growth rates; Brucella abortus; protein sequence analysis; two-dimensional gel electrophoresis
• ISSN: 1229-845X; 1976-555X; 1976-555X
• DOI: 10.4142/jvs.2018.19.2.216

is a bacterium that causes brucellosis and is the causative agent of worldwide zoonoses. Pathogenesis of the infection is complicated, and several researchers have attempted to elucidate the infection mechanism of . While several proteins have been revealed as pathogenic factors by previous researchers, the underlying mechanism of infection is unresolved. In this study, we identified proteins showing different expression levels in mutants with different biological characteristics that were generated by random insertion of a transposon. Five mutants were selected based on biological characteristics, in particular, their growth features. Total proteins of mutant and wild-type were purified and subjected to two-dimensional gel electrophoresis. Thirty protein spots of each mutant with expression increases or decreases were selected; those with a change of more than 2-fold were compared with the wild-type. Selected spots underwent liquid chromatography tandem mass spectrometry for peptide analysis. DnaK and ClpB, involved in protein aggregation, increased. SecA and GAPDH, associated with energy metabolism, decreased in some mutants with a growth rate slower than that of the wild-type. Mutants with slower growth showed a decrease in energy metabolism-related proteins, while mutants with faster growth showed an increase in pathogenicity-related proteins.