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

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

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 enzy...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 2017-07, Vol.114 (29), p.7617-7622
Hauptverfasser: Pham, Truc V., Murkin, Andrew S., Moynihan, Margaret M., Harris, Lawrence, Tyler, Peter C., Shetty, Nishant, Sacchettini, James C., Huang, Hsiao-ling, Meek, Thomas D.
Format: Artikel
Sprache:eng
Schlagworte:
2-vinyl isocitrate
Aldehydes
Bacteria
BASIC BIOLOGICAL SCIENCES
Biochemistry
Biological Sciences
Covalence
covalent adduct
Crystallography
Deactivation
Inactivation
Ionization
Isocitrate lyase
Mass spectrometry
Mass spectroscopy
mechanism-based inactivation
Mycobacterium tuberculosis
Tuberculosis
X-ray crystallography
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Zusammenfassung: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,
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1706134114