Title
Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds
Date Issued
01 December 2014
Access level
open access
Resource Type
journal article
Author(s)
Bueno C.
Santos M.
Pando-Robles V.
Batista C.V.
Johns Hopkins University
Publisher(s)
Churchill Livingstone
Abstract
Recombinant wild-pyrazinamidase from H37Rv Mycobacterium tuberculosis was analyzed by gel electrophoresis under differential reducing conditions to evaluate its quaternary structure. PZAse was fractionated by size exclusion chromatography under non-reducing conditions. PZAse activity was measured and mass spectrometry analysis was performed to determine the identity of proteins by de novo sequencing and to determine the presence of disulfide bonds. This study confirmed that M. tuberculosis wild type PZAse was able to form homo-dimers in vitro. Homo-dimers showed a slightly lower specific PZAse activity compared to monomeric PZAse. PZAse dimers were dissociated into monomers in response to reducing conditions. Mass spectrometry analysis confirmed the existence of disulfide bonds (C72-C138 and C138-C138) stabilizing the quaternary structure of the PZAse homo-dimer.
Start page
644
End page
648
Volume
94
Issue
6
Language
English
OCDE Knowledge area
Enfermedades infecciosas
Sistema respiratorio
Subjects
Scopus EID
2-s2.0-84919492367
PubMed ID
Source
Tuberculosis
ISSN of the container
1472-9792
Sponsor(s)
This research was funded by the Wellcome Trust Intermediate Fellowship awarded to PS and by the National Institute of Allergy and Infectious Diseases , National Institutes of Health US , under the terms of Award 1R01TW008669-01 awarded to MZ. D.R. was supported by an scholarship of the Franco-Peruvian School of Life Sciences.
Sources of information:
Directorio de Producción Científica
Scopus