About this item:

123 Views | 77 Downloads

Author Notes:

Correspondence: rbanerje@umich.edu

Author contributions: M.R. and G.C.C. performed the majority of the biochemical studies, H.G. did the kinetic analysis of hMCM, and L.M. and M.R. cloned and expressed the Mtb genes used in this study.

M.P. and M.K. were responsible for the crystallographic analyses, and K.W. was responsible for the EPR spectroscopic analysis. H.S. performed the macrophage experiments, and J.Z. and S.W. performed the Mtb growth experiments. M.R., G.C.C., and R.B. wrote the manuscript.

All authors were involved with data analysis of experiments performed by them or in their laboratories and edited the manuscript.

We acknowledge H. Sharma (University of Michigan) for his assistance with crystallization and the GM/CA CAT at the Advanced Light Source for beam time.

Disclosures: V.K.M. is an Investigator of the Howard Hughes Medical Institute. V.K.M. is a paid advisor to Janssen Pharmaceuticals and 5AM Ventures and owns equity in Raze Therapeutics.

Subjects:

Research Funding:

We also acknowledge the NIH Common Fund Metabolite Standards Synthesis Core (NHLBI contract no. HHSN268201300022C) for providing [13C]itaconate.

his work was supported in part by grants from the National Institutes of Health (RO1-DK45776 to R.B., 5 F32 GM113405 to G.C.C., K99-GM124296 to H.S., R35GM122455 to V.K.M., R01-DK054514 to K.W., and U19AI107774 to E.J.R.) and start-up funds from the University of Michigan (to M.K.).

Keywords:

  • Biocatalysis
  • Catalytic Domain
  • Coenzyme A
  • Crystallography, X-Ray
  • Deoxyadenosines
  • Electron Spin Resonance Spectroscopy
  • Humans
  • Hydrogen Bonding
  • Macrophages
  • Methylmalonyl-CoA Mutase
  • Models, Molecular
  • Mycobacterium tuberculosis
  • Propionates
  • Protein Conformation
  • Protein Multimerization
  • Protein Subunits
  • Succinates
  • Vitamin B 12

Itaconyl-CoA forms a stable biradical in methylmalonyl-CoA mutase and derails its activity and repair

Show all authors Show less authors

Tools:

Journal Title:

Science

Volume:

Volume 366, Number 6465

Publisher:

, Pages 589-593

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Itaconate is an immunometabolite with both anti-inflammatory and bactericidal effects. Its coenzyme A (CoA) derivative, itaconyl-CoA, inhibits B12-dependent methylmalonyl-CoA mutase (MCM) by an unknown mechanism. We demonstrate that itaconyl-CoA is a suicide inactivator of human and Mycobacterium tuberculosis MCM, which forms a markedly air-stable biradical adduct with the 5′-deoxyadenosyl moiety of the B12 coenzyme. Termination of the catalytic cycle in this way impairs communication between MCM and its auxiliary repair proteins. Crystallography and spectroscopy of the inhibited enzyme are consistent with a metal-centered cobalt radical ~6 angstroms away from the tertiary carbon-centered radical and suggest a means of controlling radical trajectories during MCM catalysis. Mycobacterial MCM thus joins enzymes in the glyoxylate shunt and the methylcitrate cycle as targets of itaconate in pathogen propionate metabolism.

Copyright information:

© 2019 American Association for the Advancement of Science. All rights reserved.

Export to EndNote