Membrane topology of the high-affinity L-glutamate transporter (GLAST-1) of the central nervous system
| Autor: | Wilhelm Stoffel, Stephan Wahle |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Central Nervous System
Neurotransmitter transporter Cytoplasm Glycosylation Amino Acid Transport System X-AG Recombinant Fusion Proteins Xenopus Molecular Sequence Data Protein Structure Secondary Cell membrane Complementary DNA medicine Animals Amino Acid Sequence Peptide sequence Conserved Sequence Glycoproteins biology Cell Membrane Biological Transport Transporter Articles Cell Biology biology.organism_classification Transmembrane protein Protein Structure Tertiary Cell biology medicine.anatomical_structure Biochemistry Membrane topology Oocytes Female Carrier Proteins |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| DOI: | 10.1083/jcb.135.6.1867 |
| Popis: | The membrane topology of the high affinity, Na(+)-coupled L-glutamate/L-aspartate transporter (GLAST-1) of the central nervous system has been determined. Truncated GLAST-1 cDNA constructs encoding protein fragments with an increasing number of hydrophobic regions were fused to a cDNA encoding a reporter peptide with two N-glycosylation sites. The respective cRNA chimeras were translated in vitro and in vivo in Xenopus oocytes. Posttranslational N-glycosylation of the two reporter consensus sites monitors the number, size, and orientation of membrane-spanning domains. The results of our experiments suggest a novel 10-transmembrane domain topology of GLAST-1, a representative of the L-glutamate neurotransmitter transporter family, with its NH2 and COOH termini on the cytoplasmic side, six NH2-terminal hydrophobic transmembrane alpha-helices, and four COOH-terminal short hydrophobic domains spanning the bilayer predicted as beta-sheets. |
| Databáze: | OpenAIRE |
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