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J Vet Sci 2018; 19(2): 216-231  https://doi.org/10.4142/jvs.2018.19.2.216
Analysis of protein expression in Brucella abortus mutants with different growth rates by two-dimensional gel electrophoresis and LC-MS/MS peptide analysis
Woo Bin Park1, Young Bin Im1, Soojin Shim1, Han Sang Yoo1,2,*
1Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea
2Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea
Correspondence to: Tel: +82-2-880-1263; Fax: +82-2-874-2738; E-mail: yoohs@snu.ac.kr
Received: May 18, 2017; Revised: September 12, 2017; Accepted: September 22, 2017; Published online: March 31, 2018.
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.
Keywords: Brucella abortus, growth rates, protein sequence analysis, two-dimensional gel electrophoresis


© 2018 The Korean Society of Veterinary Science.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.