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J Biol Chem, Vol. 275, Issue 18, 13259-13265, May 5, 2000
From the A high affinity glutathione transporter has been
identified, cloned, and characterized from the yeast
Saccharomyces cerevisiae. This transporter, Hgt1p,
represents the first high affinity glutathione transporter to be
described from any system so far. The strategy for the identification
involved investigating candidate glutathione transporters from the
yeast genome sequence project followed by genetic and physiological
investigations. This approach revealed HGT1 (open reading
frame YJL212c) as encoding a high affinity glutathione
transporter. Yeast strains deleted in HGT1 did not show any
detectable plasma membrane glutathione transport, and hgt1
Hgt1p, a High Affinity Glutathione Transporter from the Yeast
Saccharomyces cerevisiae*
§,
,, and
ESA CNRS 6161, Laboratoire de Physiologie et
Biochimie Végétales, University of Poitiers, UFR
Sciences, 40 Avenue du Recteur Pineau, 86022 Poitiers Cédex,
France and the ¶ Institute of Microbial Technology, Sector 39-A,
Chandigarh 160 036, India
disruptants were non-viable in a glutathione
biosynthetic mutant (gsh1) background. The glutathione
repressible transport activity observed in wild type cells was also
absent in the hgt1 strains. The transporter was cloned
and kinetic studies indicated that Hgt1p had a high affinity for
glutathione (Km = 54 µM)) and was not
sensitive to competition by amino acids, dipeptides, or other
tripeptides. Significant inhibition was observed, however, with
oxidized glutathione and glutathione conjugates. The transporter reveals a novel class of transporters that has homologues in other yeasts and plants but with no apparent homologues in either
Escherichia coli or in higher eukaryotes other than plants.
*
This work was supported by the Department of Science and
Technology, Government of India, the Council of Scientific and
Industrial Research (India), and the Center National de la Recherche
Scientifique (France).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.
Junior Research Fellow of the Council of Scientific and
Industrial Research, India.
**
Present address: MGH Cancer Center, Massachusetts General Hospital,
Charlestown, MA 02129.
To whom correspondence should be addressed. Tel.:
91-172-690908; Fax: 91-172-690585 or 91-172-690632; E-mail:
abachhawat@excite.com.
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