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

作者：Rueda Daniel; Sheen Patricia; Gilman Robert H; Bueno Carlos; Santos Marco; Pando Robles Victoria; Batista Cesar V; Zimic Mirko^*

来源：Tuberculosis, 2014, 94(6): 644-648.

DOI：10.1016/j.tube.2014.08.008

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

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. %26lt;br%26gt;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.

• 出版日期2014-12