期刊
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
卷 62, 期 9, 页码 -出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/AAC.00366-18
关键词
mcr-3; mcr-1; colistin resistance; homology modeling; phosphatidylethanolamine
资金
- National Natural Science Foundation of China [31602107, 81661138002]
- Capital's Funds for Health Improvement and Research [2018-4-3017]
- Medical Research Council grant DETER-XDR-CHINA [MR/P007295/1]
- BBSRC [BB/R012776/1] Funding Source: UKRI
- MRC [MR/N028317/1, MR/P007295/1] Funding Source: UKRI
The global emergence of plasmid-mediated colistin resistance genes mcr-1 and mcr-3 has threatened the role of the last-resort drug colistin in the defense against infections caused by multidrug-resistant Gram-negative bacteria. However, functional differences between these two genes in mediating colistin resistance remain poorly understood. Protein sequence alignment of MCR-3 and MCR-1 was therefore conducted in Clustal Omega to identify sequence divergence. The molecular recognition of lipid A head group phosphatidylethanolamine and MCR-3 enzyme was studied by homology modeling and molecular docking, with the catalytic mechanism of MCR-3 also being explored. Thr277 in MCR-3 was validated as the key amino acid residue responsible for the catalytic reaction using site-directed mutagenesis and was shown to act as a nucleophile. Lipid A modification induced by the MCR-3 and MCR-1 enzymes was confirmed by electrospray ionization-time of flight mass spectrometry. Far-UV circular dichroism spectra of the MCR-3 and MCR-1 enzymes suggested that MCR-3 was more thermostable than MCR-1, with a melting temperature of 66.19 degrees C compared with 61.14 degrees C for MCR-1. These data provided molecular insight into the functional differences between mcr-3 and mcr-1 in conferring colistin resistance.
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