Antibiotic resistance in Mycobacterium tuberculosis: peroxidase intermediate bypass causes poor isoniazid activation by the S315G mutant of M. tuberculosis catalase-peroxidase (KatG)
- PMID: 19363028
- PMCID: PMC2713512
- DOI: 10.1074/jbc.M109.005546
Antibiotic resistance in Mycobacterium tuberculosis: peroxidase intermediate bypass causes poor isoniazid activation by the S315G mutant of M. tuberculosis catalase-peroxidase (KatG)
Abstract
KatG (catalase-peroxidase) in Mycobacterium tuberculosis is responsible for activation of isoniazid (INH), a pro-drug used to treat tuberculosis infections. Resistance to INH is a global health problem most often associated with mutations in the katG gene. The origin of INH resistance caused by the KatG[S315G] mutant enzyme is examined here. Overexpressed KatG[S315G] was characterized by optical, EPR, and resonance Raman spectroscopy and by studies of the INH activation mechanism in vitro. Catalase activity and peroxidase activity with artificial substrates were moderately reduced (50 and 35%, respectively), whereas the rates of formation of oxyferryl heme:porphyrin pi-cation radical and the decay of heme intermediates were approximately 2-fold faster in KatG[S315G] compared with WT enzyme. The INH binding affinity for the resting enzyme was unchanged, whereas INH activation, measured by the rate of formation of an acyl-nicotinamide adenine dinucleotide adduct considered to be a bactericidal molecule, was reduced by 30% compared with WT KatG. INH resistance is suggested to arise from a redirection of catalytic intermediates into nonproductive reactions that interfere with oxidation of INH. In the resting mutant enzyme, a rapid evolution of 5-c heme to 6-c species occurred in contrast with the behavior of WT KatG and KatG[S315T] and consistent with greater flexibility at the heme edge in the absence of the hydroxyl of residue 315. Insights into the effects of mutations at residue 315 on enzyme structure, peroxidation kinetics, and specific interactions with INH are presented.
Figures








Similar articles
-
Isoniazid-resistance conferring mutations in Mycobacterium tuberculosis KatG: catalase, peroxidase, and INH-NADH adduct formation activities.Protein Sci. 2010 Mar;19(3):458-74. doi: 10.1002/pro.324. Protein Sci. 2010. PMID: 20054829 Free PMC article.
-
Reduced affinity for Isoniazid in the S315T mutant of Mycobacterium tuberculosis KatG is a key factor in antibiotic resistance.J Biol Chem. 2003 Apr 25;278(17):14769-75. doi: 10.1074/jbc.M300326200. Epub 2003 Feb 13. J Biol Chem. 2003. PMID: 12586821
-
Analysis of interactions of clinical mutants of catalase-peroxidase (KatG) responsible for isoniazid resistance in Mycobacterium tuberculosis with derivatives of isoniazid.J Glob Antimicrob Resist. 2017 Dec;11:57-67. doi: 10.1016/j.jgar.2017.06.014. Epub 2017 Jul 23. J Glob Antimicrob Resist. 2017. PMID: 28743650
-
Resistance to Isoniazid and Ethionamide in Mycobacterium tuberculosis: Genes, Mutations, and Causalities.Microbiol Spectr. 2014 Aug;2(4):MGM2-0014-2013. doi: 10.1128/microbiolspec.MGM2-0014-2013. Microbiol Spectr. 2014. PMID: 26104204 Free PMC article. Review.
-
Impact of isoniazid resistance on virulence of global and south Indian clinical isolates of Mycobacterium tuberculosis.Tuberculosis (Edinb). 2014 Dec;94(6):557-63. doi: 10.1016/j.tube.2014.08.011. Epub 2014 Sep 6. Tuberculosis (Edinb). 2014. PMID: 25270728 Review.
Cited by
-
Modeling the structural origins of drug resistance to isoniazid via key mutations in Mycobacterium tuberculosis catalase-peroxidase, KatG.Tuberculosis (Edinb). 2018 Jan;108:155-162. doi: 10.1016/j.tube.2017.11.007. Epub 2017 Nov 22. Tuberculosis (Edinb). 2018. PMID: 29523317 Free PMC article.
-
Mutations in catalase-peroxidase KatG from isoniazid resistant Mycobacterium tuberculosis clinical isolates: insights from molecular dynamics simulations.J Mol Model. 2017 Apr;23(4):121. doi: 10.1007/s00894-017-3290-3. Epub 2017 Mar 16. J Mol Model. 2017. PMID: 28303436
-
Is IQG-607 a Potential Metallodrug or Metallopro-Drug With a Defined Molecular Target in Mycobacterium tuberculosis?Front Microbiol. 2018 May 1;9:880. doi: 10.3389/fmicb.2018.00880. eCollection 2018. Front Microbiol. 2018. PMID: 29765372 Free PMC article. Review.
-
Isoniazid and thioacetazone may exhibit antitubercular activity by binding directly with the active site of mycolic acid cyclopropane synthase: Hypothesis based on computational analysis.Bioinformation. 2012;8(16):787-9. doi: 10.6026/97320630008787. Epub 2012 Aug 24. Bioinformation. 2012. PMID: 23055630 Free PMC article.
-
Isoniazid-resistance conferring mutations in Mycobacterium tuberculosis KatG: catalase, peroxidase, and INH-NADH adduct formation activities.Protein Sci. 2010 Mar;19(3):458-74. doi: 10.1002/pro.324. Protein Sci. 2010. PMID: 20054829 Free PMC article.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Medical
Research Materials
Miscellaneous