Analysis of outer membrane proteome of Escherichia coli related to resistance to ampicillin and tetracycline.
The elucidation of the molecular details of antibiotic resistance will lead to improvements in extending the efficacy of current antimicrobials. In the current study, proteomic methodologies were applied to characterize functional outer membrane proteins (Omps) of E. coli K-12 responded to tetracycline and ampicillin resistance for understanding of universal pathways that form barriers for antimicrobial agents. For this purpose, E. coli K-12 expressional outer membrane proteome was characterized and identified with the use of 2-DE and MALDI-TOF/MS methods. Then, differential Omps due to tetracycline or ampcilin resistance were determined by comparison between tetracycline minimum inhibitory concentration (MIC)10, ampicillin MIC10, control0 and control10, showing 9 proteins with 11 spots for tetracycline and 8 protein with 9 spots for ampicillin, showing a difference in only 1 protein (decreased LamB in tetracyclin) between the two antibiotics. Among the proteins, 3 were known as antibiotic-resistant proteins, including TolC, OmpC and YhiU, while FimD precursor, LamB, Tsx, YfiO, OmpW, NlpB were first reported here to be antibiotic-resistance-related proteins. Our findings will be helpful for further understanding of antibiotic-resistant mechanism(s). This study also shows that the combination of Omp purification methods certainly contributes the sensitivity of Omp detection.
Xu
Changxin
C
Center for Proteomics, Department of Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian, PR China.
Lin
Xiangmin
X
Ren
Haixia
H
Zhang
Yueling
Y
Wang
Sanying
S
Peng
Xuanxian
X
D000667:Ampicillin / Q000494:pharmacology*; D000668:Ampicillin Resistance; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D048168:Escherichia coli K12 / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013753:Tetracycline Resistance; D013754:Tetracyclines / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D020543:Proteome; D013754:Tetracyclines; D000667:Ampicillin