Relationship between the mutation ACT271CCT (Thr271Pro) in katG gene of Mycobacterium tuberculosis and the drug resistance of isoniazid
YANG Shou-hui1, LIU Wei-you2, WU Long-huo3, YUAN Xiao-liang2
1. Gannan Medical University, Ganzhou 341000, China; 2.Department of Respiratory Medicine, the First Affiliated Hospital of Gannan Medical University, Ganzhou 341000, China; 3. College of Pharmacy, Gannan Medical University, Ganzhou 341000, China
Abstract:To explore the effect of ACT271CCT (Thr271Pro) mutation in katG gene of Mycobacterium tuberculosis on isoniazid resistance, wild-type katG, mutant (AGC315ACC) katG and (ACT271CCT) katG gene were artificially synthesized. After these target genes were cloned into pET22b(+) plasmids, the recombinant plasmids were transferred into an expression host, Escherichia coli BL21 (DE3) pLySs. The expression of recombinant proteins was induced by the addition of IPTG. The target proteins were purified using affinity chromatography, and the peroxidase and catalase activity of recombinant katG proteins were detected. In addition, the semi-flexible docking mode was used to simulate the interaction between wild-type or Thr271Pro mutant katG protein and isoniazid using software AutoDock4.0. Results showed that the recombinant katG proteins were purified successfully. Compared with wild-type katG protein, the catalase and peroxidase activity of Thr271Pro mutant katG protein decreased by 56% and 63% (158.8 U/mg and 28.7 U/mg) respectively, while those of Ser315Thr mutant katG proteins decreased by 74% and 75% (96.2 U/mg and 19.7 U/mg) respectively. The molecular docking results showed that the binding energy of isoniazid and Thr271Pro mutant katG protein increased, and the amino acid residues of Thr271Pro mutant led to a certain deformation of katG protein oxidative reactive cavity in comparison with the wild-type. Thr271Pro mutation in KatG gene results in partial loss of katG enzyme activity and deformation of katG protein oxidative reactive cavity, and might confer resistance to isoniazid.
杨寿慧, 刘惟优, 吴龙火, 袁小亮 . 结核分枝杆菌KatG-ACT271CCT(Thr271Pro)突变与异烟肼耐药关系的研究[J]. 中国人兽共患病学报, 2019, 35(6): 487-492.
YANG Shou-hui, LIU Wei-you, WU Long-huo, YUAN Xiao-liang. Relationship between the mutation ACT271CCT (Thr271Pro) in katG gene of Mycobacterium tuberculosis and the drug resistance of isoniazid. Chinese Journal of Zoonoses, 2019, 35(6): 487-492.
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