I. Choi et Sy. Chiu, Loss of cell viability by histidine substitution of leucine 325 of the glutamate transporter EAAT1, BIOC BIOP R, 275(2), 2000, pp. 382-385
Citations number
24
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Although glutamate transporters and neutral amino acid transporters have 55
% amino acid identity in the transmembrane domains, many residues are still
unique to individual transporters, providing for structural stability or s
ubstrate binding. In this study, the mutant protein L325H, which replaced a
leucine 325 of the glutamate transporter EAAT1 by a histidine, was evaluat
ed. When expressed in Xenopus oocytes, L325H caused oocytes to weaken pigme
ntation in the animal pole, accompanied by patches of colorless spots. Oocy
tes finally oozed cytoplasm. The resting membrane potential in L325H oocyte
s was -18.9 +/- 2.5 mV, significantly more positive than -37.3 +/- 2.5 mV o
f oocytes expressing EAAT1. The holding current at -60 mV was 283.1 +/- 48.
3 nA in L325H oocytes and 92.2 +/- 12.6 nA in EAAT1 oocytes. These results
suggest that even though glutamate and neutral amino acid transporters have
strong overall homology, the local structure in the transmembrane domains
may be different. (C) 2000 Academic Press.