Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds

Daniel Rueda; Patricia Sheen; Robert H. Gilman; Carlos Bueno; Marco Santos; Victoria Pando-Robles; Cesar V. Batista; Mirko Zimic

doi:10.1016/j.tube.2014.08.008

Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds

Daniel Rueda
, Patricia Sheen
, Robert H. Gilman
, Carlos Bueno
, Marco Santos
, Victoria Pando-Robles
, Cesar V. Batista
, Mirko Zimic

School of Science and Engineering

Universidad Peruana Cayetano Heredia
Johns Hopkins Bloomberg School of Public Health
National Institute of Public Health
UNAM

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

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.

Original language	English
Pages (from-to)	644-648
Number of pages	5
Journal	Tuberculosis
Volume	94
Issue number	6
DOIs	https://doi.org/10.1016/j.tube.2014.08.008
State	Published - 1 Dec 2014

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Dimer
Drug resistance
Multimer
Mycobacterium
Nicotinamidase
Nicotinamide
Pyrazinamidase
Pyrazinamide
Reducing stress
Tuberculosis

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10.1016/j.tube.2014.08.008

Cite this

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title = "Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds",

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.",

keywords = "Dimer, Drug resistance, Multimer, Mycobacterium, Nicotinamidase, Nicotinamide, Pyrazinamidase, Pyrazinamide, Reducing stress, Tuberculosis",

author = "Daniel Rueda and Patricia Sheen and Gilman, \{Robert H.\} and Carlos Bueno and Marco Santos and Victoria Pando-Robles and Batista, \{Cesar V.\} and Mirko Zimic",

note = "Publisher Copyright: {\textcopyright} 2014 Elsevier Ltd.",

year = "2014",

month = dec,

day = "1",

doi = "10.1016/j.tube.2014.08.008",

language = "Ingl{\'e}s",

volume = "94",

pages = "644--648",

journal = "Tuberculosis",

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TY - JOUR

T1 - Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds

AU - Rueda, Daniel

AU - Sheen, Patricia

AU - Gilman, Robert H.

AU - Bueno, Carlos

AU - Santos, Marco

AU - Pando-Robles, Victoria

AU - Batista, Cesar V.

AU - Zimic, Mirko

PY - 2014/12/1

Y1 - 2014/12/1

N2 - 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.

AB - 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.

KW - Dimer

KW - Drug resistance

KW - Multimer

KW - Mycobacterium

KW - Nicotinamidase

KW - Nicotinamide

KW - Pyrazinamidase

KW - Pyrazinamide

KW - Reducing stress

KW - Tuberculosis

UR - https://www.scopus.com/pages/publications/84919492367

U2 - 10.1016/j.tube.2014.08.008

DO - 10.1016/j.tube.2014.08.008

M3 - Artículo

C2 - 25199451

AN - SCOPUS:84919492367

SN - 1472-9792

VL - 94

SP - 644

EP - 648

JO - Tuberculosis

JF - Tuberculosis

IS - 6

ER -

Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds

Abstract

UN SDGs

Keywords

Access to Document

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