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J. Biol. Chem., Vol. 276, Issue 6, 3798-3804, February 9, 2001
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From the Harold C. Simmons Arthritis Research Center, Department of
Internal Medicine, University of Texas Southwestern Medical Center at
Dallas, Dallas, Texas 75390
The ectoenzyme,
Expression of
-Glutamyl Transpeptidase Protects Ramos B
Cells from Oxidation-induced Cell Death*
,
-glutamyl transpeptidase (GGT,
EC 2.3.2.2) cleaves glutathione (GSH) to facilitate the recapture of
cysteine for synthesis of intracellular GSH. The impact of GGT
expression on cell survival during oxidative stress was investigated
using the human B cell lymphoblastoid cell line, Ramos. Ramos cells did
not express surface GGT and exhibited no GGT enzyme activity. In
contrast, Ramos cells stably transfected with the human GGT cDNA
expressed high levels of surface GGT and enzymatic activity. GGT-transfected Ramos cells were protected from apoptosis when cultured
in cyst(e)ine-deficient medium. The GGT-expressing cells also had lower
levels of intracellular reactive oxygen species (ROS). Homocysteic acid
and alanine, inhibitors of cystine and cysteine uptake, respectively,
caused increased ROS content and diminished viability of GGT expressing
cells. Exogenous GSH increased the viability of the GGT-transfected
cells more effectively than that of control cells, whereas the products
of GSH metabolism prevented death of both the control and
GGT-transfected cells comparably. These data indicate that GGT cleavage
of GSH and the subsequent recapture of cysteine and cystine allow cells
to maintain low levels of cellular ROS and thereby avoid apoptosis
induced by oxidative stress.
*
This work was supported in part by United States Public
Health Service Grant AI42772 (to D. R. K.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Simmons Arthritis
Research Center, University of Texas Southwestern Medical Center at
Dallas, 5323 Harry Hines Blvd., Dallas, TX 75390-8884. E-mail: David.Karp@UTSouthwestern.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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