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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, Shim Soojin1, 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: October 13, 2017.
Brucella abortus is a bacterium that causes brucellosis and is the causative agent of worldwide zoonosis. Pathogenesis of the Brucella abortus infection is so complicates. Thus, several researches have made to solve mechanism in infection of Brucella abortus. While several proteins are revealed as the pathogenic factors by previous researches, underlying mechanism of the Brucella abortus infection is still left to be resolved. We identify the proteins showing the different expression level in Brucella abortus mutants with different biological characteristics that generated by random insertion of transposon. Five mutants were selected based on the biological characteristics, especially the growth features. Total proteins of mutants and wild type were purified and subjected to two dimensional gel electrophoresis. 30 spots of proteins were selected each with an increase and decrease, a change of more than 2-fold was confirmed as compared with the wild type. Selected spots were analyzed using LC-MS/MS peptide analysis. DnaK and ClpB involved in protein aggregation were simultaneously increased. Proteins associated with energy metabolism, SecA and GAPDH were reduced in the expression in some mutants that slower growth rates than wild type. Mutants with slower growth showed decrease with energy metabolism related proteins, while mutants with faster growth showed increase in the pathogenicity related proteins.
Keywords: Brucella abortus, Two dimensional gel electrophoresis, LC-MS/MS peptide analysis, Growth rates

© 2017 The Korean Society of Veterinary Science.