Analysis of protein expression in Brucella abortus mutants with different growth rates by two-dimensional gel electrophoresis and LC-MS/MS peptide analysis
10.4142/jvs.2018.19.2.216
- Author:
Woo Bin PARK
1
;
Young Bin IM
;
Soojin SHIM
;
Han Sang YOO
Author Information
1. Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea. yoohs@snu.ac.kr
- Publication Type:Original Article
- Keywords:
Brucella abortus;
growth rates;
protein sequence analysis;
two-dimensional gel electrophoresis
- MeSH:
Brucella abortus;
Brucella;
Brucellosis;
Chromatography, Liquid;
Electrophoresis, Gel, Two-Dimensional;
Energy Metabolism;
Population Characteristics;
Sequence Analysis, Protein;
Tandem Mass Spectrometry;
Zoonoses
- From:Journal of Veterinary Science
2018;19(2):216-231
- CountryRepublic of Korea
- Language:English
-
Abstract:
Brucella abortus is a bacterium that causes brucellosis and is the causative agent of worldwide zoonoses. Pathogenesis of the B. abortus infection is complicated, and several researchers have attempted to elucidate the infection mechanism of B. abortus. While several proteins have been revealed as pathogenic factors by previous researchers, the underlying mechanism of B. abortus infection is unresolved. In this study, we identified proteins showing different expression levels in B. abortus 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 B. abortus 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.
