Cooperative binding of gamma-glutamyl substrate to human glutathione synthetase.

Abstract:

:Human glutathione synthetase is responsible for catalyzing the final step in glutathione biosynthesis. It is a homodimer with a monomer subunit MW of 52 kDa. Kinetic analysis reveals a departure from linearity of the Lineweaver-Burk double reciprocal plot for the binding of gamma-glutamyl substrate, indicating cooperative binding. The measured apparent K(m) values for gamma-glutamyl-alpha-aminobutyrate (an analog of gamma-glutamyl-alpha-aminobutyrate) are 63 and 164 microM, respectively. Neither ATP (K(m) of 248 microM) nor glycine (K(m) of 452 microM) exhibits such cooperative binding behavior. Although ATP is proposed to play a key role in the sequential binding of gamma-glutamyl substrate to the enzyme, the cooperative binding of the gamma-glutamyl substrate is not affected by alterations of ATP concentration. Quantitative analysis of the kinetic results for gamma-glutamyl substrate binding gives a Hill coefficient (h) of 0.75, indicating negative cooperativity. Our studies, for the first time, show that human glutathione synthetase is an allosteric enzyme with cooperative binding for gamma-glutamyl substrate.

authors

Njalsson R,Norgren S,Larsson A,Huang CS,Anderson ME,Luo JL

doi

10.1006/bbrc.2001.5961

subject

Has Abstract

pub_date

2001-11-23 00:00:00

pages

80-4

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(01)95961-3

journal_volume

289

pub_type

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