Unknown

Dataset Information

0

Mechanism-based inactivator of isocitrate lyases 1 and 2 from Mycobacterium tuberculosis.

ABSTRACT: Isocitrate lyase (ICL, types 1 and 2) is the first enzyme of the glyoxylate shunt, an essential pathway for Mycobacterium tuberculosis (Mtb) during the persistent phase of human TB infection. Here, we report 2-vinyl-d-isocitrate (2-VIC) as a mechanism-based inactivator of Mtb ICL1 and ICL2. The enzyme-catalyzed retro-aldol cleavage of 2-VIC unmasks a Michael substrate, 2-vinylglyoxylate, which then forms a slowly reversible, covalent adduct with the thiolate form of active-site Cys191 2-VIC displayed kinetic properties consistent with covalent, mechanism-based inactivation of ICL1 and ICL2 with high efficiency (partition ratio, <1). Analysis of a complex of ICL1:2-VIC by electrospray ionization mass spectrometry and X-ray crystallography confirmed the formation of the predicted covalent S-homopyruvoyl adduct of the active-site Cys191.

SUBMITTER: Pham TV 

PROVIDER: S-EPMC5530696 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

Json Xml
altmetric image

Publications

Mechanism-based inactivator of isocitrate lyases 1 and 2 from <i>Mycobacterium tuberculosis</i>.

Pham Truc V TV   Murkin Andrew S AS   Moynihan Margaret M MM   Harris Lawrence L   Tyler Peter C PC   Shetty Nishant N   Sacchettini James C JC   Huang Hsiao-Ling HL   Meek Thomas D TD  

Proceedings of the National Academy of Sciences of the United States of America 20170705 29

Isocitrate lyase (ICL, types 1 and 2) is the first enzyme of the glyoxylate shunt, an essential pathway for <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) during the persistent phase of human TB infection. Here, we report 2-vinyl-d-isocitrate (2-VIC) as a mechanism-based inactivator of <i>Mtb</i> ICL1 and ICL2. The enzyme-catalyzed retro-aldol cleavage of 2-VIC unmasks a Michael substrate, 2-vinylglyoxylate, which then forms a slowly reversible, covalent adduct with the thiolate form of active-s  ...[more]

Similar Datasets

Unknown Mycobacterium tuberculosis isocitrate lyases 1 and 2 are jointly required for in vivo growth and virulence.
| S-EPMC1464426 | biostudies-literature
Unknown Structural, kinetic and chemical mechanism of isocitrate dehydrogenase-1 from Mycobacterium tuberculosis.
| S-EPMC3706558 | biostudies-literature
Unknown Demystifying the catalytic pathway of Mycobacterium tuberculosis isocitrate lyase.
| S-EPMC7609661 | biostudies-literature
Unknown Acetyl-CoA-mediated activation of Mycobacterium tuberculosis isocitrate lyase 2.
| S-EPMC6788997 | biostudies-literature
Unknown Potential inhibitors for isocitrate lyase of Mycobacterium tuberculosis and non-M. tuberculosis: a summary.
| S-EPMC4306415 | biostudies-literature
Unknown Modulation of Central Carbon Metabolism by Acetylation of Isocitrate Lyase in Mycobacterium tuberculosis.
| S-EPMC5359664 | biostudies-literature
Unknown Discovery of a Novel Inhibitor Structure of <i>Mycobacterium tuberculosis</i> Isocitrate Lyase.
| S-EPMC9026967 | biostudies-literature
Unknown Itaconate is a covalent inhibitor of the <i>Mycobacterium tuberculosis</i> isocitrate lyase.
| S-EPMC8130629 | biostudies-literature
Unknown Localization and targeting of isocitrate lyases in Saccharomyces cerevisiae.
| S-EPMC1217762 | biostudies-other
Unknown Identification of mannich base as a novel inhibitor of Mycobacterium tuberculosis isocitrate by high-throughput screening.
| S-EPMC3076504 | biostudies-literature