Cloning and expression of a rat brain GABA transporter
Abstract
A complementary DNA clone (designated GAT-1) encoding a transporter for the neurotransmitter gamma-aminobutyric acid (GABA) has been isolated from rat brain, and its functional properties have been examined in Xenopus oocytes. Oocytes injected with GAT-1 synthetic messenger RNA accumulated [^3H]GABA to levels above control values. The transporter encoded by GAT-1 has a high affinity for GABA, is sodium-and chloride-dependent, and is pharmacologically similar to neuronal GABA transporters. The GAT-1 protein shares antigenic determinants with a native rat brain GABA transporter. The nucleotide sequence of GAT-1 predicts a protein of 599 amino acids with a molecular weight of 67 kilodaltons. Hydropathy analysis of the deduced protein suggests multiple transmembrane regions, a feature shared by several cloned transporters; however, database searches indicate that GAT-1 is not homologous to any previously identified proteins. Therefore, GAT-1 appears to be a member of a previously uncharacterized family of transport molecules.
Additional Information
© 1990 American Association for the Advancement of Science. 30 March 1990; Accepted 8 June 1990. We thank R. Radian and A. Bendahan for protein purification, and I. Ahmed, J. Hoger, D. Krafte, C. LaBarca, T. Snutch, and A. Walter for sharing protocols and expertise. We also thank T. Snutch for participating in the construction of the cDNA library, S. Celniker and the Caltech fly group for the use of their oligonucleotide synthesizer, and S. Bajjalieh for comments on the manuscript. We especially thank A. Gouin for oocyte preparation. Supported by NIH grants GM 29836, GM 10991, and NS 16708, by U.S.-Israel Binational Science Foundation grant 86-00147, and by an NIH postdoctoral fellowship to J.G.Additional details
- Eprint ID
- 53114
- DOI
- 10.1126/science.1975955
- Resolver ID
- CaltechAUTHORS:20141222-150126708
- GM 29836
- NIH
- GM 10991
- NIH
- NS 16708
- NIH
- 86-00147
- Binational Science Foundation (USA-Israel)
- NIH Postdoctoral Fellowship
- Created
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2014-12-23Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field