Outer membrane proteomics of kanamycin-resistant Escherichia coli identified MipA as a novel antibiotic resistance-related protein.
Antibiotic-resistant bacteria are a great threat to human health and food safety and there is an urgent need to understand the mechanisms of resistance for combating these bacteria. In the current study, comparative proteomic methodologies were applied to identify Escherichia coli K-12 outer membrane (OM) proteins related to kanamycin resistance. Mass spectrometry and western blotting results revealed that OM proteins TolC, Tsx and OstA were up-regulated, whereas MipA, OmpA, FadL and OmpW were down-regulated in kanamycin-resistant E. coli K-12 strain. Genetic deletion of tolC (ΔtolC-Km) led to a 2-fold decrease in the minimum inhibitory concentration (MIC) of kanamycin and deletion of mipA (ΔmipA-Km) resulted in a 4-fold increase in the MIC of kanamycin. Changes in the MICs for genetically modified strains could be completely recovered by gene complementation. Compared with the wild-type strain, the survival capability of ΔompA-Km was significantly increased and that of Δtsx-Km was significantly decreased. We further evaluated the role and expression of MipA in response to four other antibiotics including nalidixic acid, streptomycin, chloramphenicol and aureomycin, which suggested that MipA was a novel OM protein related to antibiotic resistance.
Center for Proteomics and Metabolomics, State Key Laboratory of Biocontrol, MOE Key Lab Aquatic Food Safety, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China.
Hui
H
Li
School of Biological Science and Biotechnology, Minnan Normal University, Zhangzhou 363000, People's Republic of China.
Dan-feng
DF
Zhang
Agroecological Institute, Fujian Agricultural and Forestry University, Fuzhou 350002, People's Republic of China.
Xiang-min
XM
Lin
Center for Proteomics and Metabolomics, State Key Laboratory of Biocontrol, MOE Key Lab Aquatic Food Safety, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China pxuanx@sysu.edu.cn wangpeng@xmu.edu.cn.
Xuan-xian
XX
Peng
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D015153:Blotting, Western; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D048168:Escherichia coli K12 / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D007612:Kanamycin / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D040901:Proteomics
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C000604652:MipA protein, E coli; D007612:Kanamycin
  E. coli K-12; MipA; OM proteins; kanamycin resistance; proteomics