Open AccessOverexpression, purification, crystallization and preliminary X‐ray analysis of Rv2780 from Mycobacterium tuberculosis H37Rv
Author(s) -
Tripathi Sarvind Mani,
Ramachandran Ravishankar
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108007653
Subject(s) - crystallization , mycobacterium tuberculosis , tuberculosis , microbiology and biotechnology , materials science , chemistry , biology , medicine , pathology , organic chemistry
Rv2780, an alanine dehydrogenase from Mycobacterium tuberculosis (MtAlaDH), catalyzes the NAD‐dependent interconversion of alanine and pyruvate. Alanine dehydrogenase is released into the culture medium in substantial amounts by virulent strains of mycobacteria and is not found in the vaccine strain of tuberculosis. Crystals of recombinant MtAlaDH were grown from 2 M ammonium sulfate solution at ∼12 mg ml −1 protein concentration in two crystal forms which occur in the presence and absence of NAD/pyruvate, respectively. Diffraction data extending to 2.6 Å were collected at room temperature from both apo and ternary complex crystals. Crystals of the apoenzyme have unit‐cell parameters a = 173.89, b = 127.07, c = 135.95 Å. They are rod‐like in shape and belong to space group C 2. They contain a hexamer in the asymmetric unit. Crystals of the ternary complex belong to space group P 4 3 2 1 2 and have unit‐cell parameters a = b = 88.99, c = 373.85 Å. There are three subunits in the asymmetric unit of the holoenzyme crystals.