Role of metal ions on the activity of Mycobacterium tuberculosis pyrazinamidase

Olivera, P.; Christiansen, G.; Evangelista, W.; Ferrer, P.; Fuentes, P.; Gilman, Robert Hugh; Gutiérrez, A.H.; Lamba, D.; Sheen Cortavarria, Patricia; Zimic-Peralta, Mirko Juan; Rueda, D.; Solis, J.; Sotelo, J.; Moreno-Román, P.; Flores, M.; Pesaresi, A.

dc.contributor.author	Sheen Cortavarria, Patricia
dc.contributor.author	Ferrer, P.
dc.contributor.author	Gilman, Robert Hugh
dc.contributor.author	Christiansen, G.
dc.contributor.author	Moreno-Román, P.
dc.contributor.author	Gutiérrez, A.H.
dc.contributor.author	Sotelo, J.
dc.contributor.author	Evangelista, W.
dc.contributor.author	Fuentes, P.
dc.contributor.author	Rueda, D.
dc.contributor.author	Flores, M.
dc.contributor.author	Olivera, P.
dc.contributor.author	Solis, J.
dc.contributor.author	Pesaresi, A.
dc.contributor.author	Lamba, D.
dc.contributor.author	Zimic-Peralta, Mirko Juan
dc.date.accessioned	2022-01-18T19:35:51Z
dc.date.available	2022-01-18T19:35:51Z
dc.date.issued	2012
dc.identifier.uri	https://hdl.handle.net/20.500.12866/11215
dc.description.abstract	Pyrazinamidase of Mycobacterium tuberculosis catalyzes the conversion of pyrazinamide to the active molecule pyrazinoic acid. Reduction of pyrazinamidase activity results in a level of pyrazinamide resistance. Previous studies have suggested that pyrazinamidase has a metal-binding site and that a divalent metal cofactor is required for activity. To determine the effect of divalent metals on the pyrazinamidase, the recombinant wild-type pyrazinamidase corresponding to the H37Rv pyrazinamide-susceptible reference strain was expressed in Escherichia coli with and without a carboxy terminal. His-tagged pyrazinamidase was inactivated by metal depletion and reactivated by titration with divalent metals. Although Co2+, Mn2+, and Zn2+ restored pyrazinamidase activity, only Co2+ enhanced the enzymatic activity to levels higher than the wild-type pyrazinamidase. Cu2+, Fe 2+, Fe3+, and Mg2+ did not restore the activity under the conditions tested. Various recombinant mutated pyrazinamidases with appropriate folding but different enzymatic activities showed a differential pattern of recovered activity. X-ray fluorescence and atomic absorbance spectroscopy showed that recombinant wild-type pyrazinamidase expressed in E. coli most likely contained Zn. In conclusion, this study suggests that M. tuberculosis pyrazinamidase is a metalloenzyme that is able to coordinate several ions, but in vivo, it is more likely to coordinate Zn2+. However, in vitro, the metal-depleted enzyme could be reactivated by several divalent metals with higher efficiency than Zn.	en_US
dc.language.iso	eng
dc.publisher	American Society of Tropical Medicine and Hygiene
dc.relation.ispartofseries	American Journal of Tropical Medicine and Hygiene
dc.rights	info:eu-repo/semantics/restrictedAccess
dc.subject	Controlled study	en_US
dc.subject	in vitro study	en_US
dc.subject	in vivo study	en_US
dc.subject	non protein expression	en_US
dc.subject	unclassified drug	en_US
dc.subject	Mycobacterium tuberculosis	en_US
dc.subject	Kinetics	en_US
dc.subject	pyrazinamide	en_US
dc.subject	Amidohydrolases	en_US
dc.subject	atomic absorption spectrometry	en_US
dc.subject	bacterial enzyme	en_US
dc.subject	carboxy terminal sequence	en_US
dc.subject	Circular Dichroism	en_US
dc.subject	cobalt	en_US
dc.subject	divalent cation	en_US
dc.subject	enzyme activity	en_US
dc.subject	enzyme inactivation	en_US
dc.subject	enzyme reactivation	en_US
dc.subject	Escherichia coli	en_US
dc.subject	ferric ion	en_US
dc.subject	ferrous ion	en_US
dc.subject	magnesium ion	en_US
dc.subject	manganese	en_US
dc.subject	metal ion	en_US
dc.subject	Metals	en_US
dc.subject	Models, Molecular	en_US
dc.subject	pyrazinamidase	en_US
dc.subject	recombinant enzyme	en_US
dc.subject	Spectrophotometry, Atomic	en_US
dc.subject	titrimetry	en_US
dc.subject	wild type	en_US
dc.subject	X ray fluorescence	en_US
dc.subject	zinc ion	en_US
dc.title	Role of metal ions on the activity of Mycobacterium tuberculosis pyrazinamidase	en_US
dc.type	info:eu-repo/semantics/article
dc.identifier.doi	https://doi.org/10.4269/ajtmh.2012.10-0565
dc.subject.ocde	https://purl.org/pe-repo/ocde/ford#3.03.06
dc.relation.issn	1476-1645