Investigation of Rifampicin Resistance Mechanisms in Brucella abortus Using MS-Driven Comparative Proteomics

被引:34
作者
Sandalakis, Vassilios [2 ,3 ]
Psaroulaki, Anna [2 ,3 ]
De Bock, Pieter-Jan [4 ,5 ]
Christidou, Athanasia [2 ]
Gevaert, Kris [4 ,5 ]
Tsiotis, Georgios [1 ]
Tselentis, Yiannis [2 ,3 ]
机构
[1] Univ Crete, Div Biochem, Dept Chem, GR-71003 Voutes, Greece
[2] Univ Crete, Sch Med, Dept Clin Bacteriol Parasitol Zoonoses & Geog Med, GR-71110 Iraklion, Greece
[3] Reg Lab Publ Hlth 71110, Iraklion, Crete, Greece
[4] VIB, Dept Med Prot Res, B-9000 Ghent, Belgium
[5] Univ Ghent, Dept Biochem, B-9000 Ghent, Belgium
关键词
Brucella abortus; comparative; proteomics; rifampicin; in vitro; mutant; resistance; MIC; MYCOBACTERIUM-TUBERCULOSIS; ANTIBIOTIC-RESISTANCE; ESCHERICHIA-COLI; GLOBAL ANALYSIS; INTRACELLULAR SURVIVAL; STAPHYLOCOCCUS-AUREUS; CANDIDATE PROTEINS; BACILLUS-SUBTILIS; RPOB MUTATIONS; RNA-POLYMERASE;
D O I
10.1021/pr201122w
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Mutations in the rpoB gene have already been shown to contribute to rifampicin resistance in many bacterial strains including Brucella species. Resistance against this antibiotic easily occurs and resistant strains have already been detected in human samples. We here present the first research project that combines proteomic, genomic, and microbiological analysis to investigate rifampicin resistance in an in vitro developed rifampicin resistant strain of Brucella abortus 2308. In silico analysis of the rpoB gene was performed and several antibiotics used in the therapy of Brucellosis were used for cross resistance testing. The proteomic profiles were examined and compared using MS-driven comparative proteomics. The resistant strain contained an already described mutation in the rpoB gene, V154F. A correlation between rifampicin resistance and reduced susceptibility on trimethoprim/sulfamethoxazole was detected by E-test and supported by the proteomics results. Using 12 836 MS/MS spectra we identified 6753 peptides corresponding to 456 proteins. The resistant strain presented 39 differentially regulated proteins most of which are involved in various metabolic pathways. Results from our research suggest that rifampicin resistance in Brucella mostly involves mutations in the rpoB gene, excitation of several metabolic processes, and perhaps the use of the already existing secretion mechanisms at a more efficient level.
引用
收藏
页码:2374 / 2385
页数:12
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