Functional characterization of a novel ArgA from Mycobacterium tuberculosis.

Abstract:

:The Mycobacterium tuberculosis gene Rv2747 encodes a novel 19-kDa ArgA that catalyzes the initial step in L-arginine biosynthesis, namely the conversion of L-glutamate to alpha-N-acetyl-L-glutamate. Initial velocity studies reveal that Rv2747 proceeds through a sequential kinetic mechanism, with K(m) values of 280 mM for L-glutamine and 150 microM for acetyl-coenzyme A and with a k(cat) value of 200 min(-1). Initial velocity studies with L-glutamate showed that even at concentrations of 600 mM, saturation was not observed. Therefore, only a k(cat)/K(m) value of 125 M(-1) min(-1) can be calculated. Inhibition studies reveal that the enzyme is strongly regulated by L-arginine, the end product of the pathway (50% inhibitory concentration, 26 microM). The enzyme was completely inhibited by 500 microM arginine, with a Hill coefficient of 0.60, indicating negatively cooperative binding of L-arginine.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Errey JC,Blanchard JS

doi

10.1128/JB.187.9.3039-3044.2005

keywords:

subject

Has Abstract

pub_date

2005-05-01 00:00:00

pages

3039-44

issue

9

eissn

0021-9193

issn

1098-5530

pii

187/9/3039

journal_volume

187

pub_type

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