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Isolation of a cDNA encoding the human brain serotonin transporter
K P Lesch1, B L Wolozin, H C Estler
1Department of Psychiatry, University of Würzburg, Federal Republic of Germany.
Journal of Neural Transmission. General Section
|January 1, 1993
Summary
Researchers isolated and sequenced the human serotonin transporter (5-HTT) cDNA from the dorsal raphe nucleus. This transporter shows high homology to the rat protein but has unique phosphorylation sites, suggesting distinct regulatory mechanisms.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- The serotonin transporter (5-HTT) plays a crucial role in regulating serotonergic neurotransmission.
- Understanding the molecular characteristics of human 5-HTT is essential for comprehending its function and potential therapeutic targets.
Purpose of the Study:
- To isolate and sequence the complementary DNA (cDNA) encoding the human serotonin transporter (5-HTT) from the dorsal raphe nucleus.
- To characterize the structural and potential regulatory features of the human 5-HTT.
Main Methods:
- Cross-species amplification of human 5-HTT partial cDNA using the polymerase chain reaction (PCR).
- Rapid amplification of 3' and 5' cDNA ends (RACE-PCR) to obtain full-length cDNA.
- Bioinformatic analysis including hydropathicity and homology comparisons.
Main Results:
- A full-length cDNA encoding human 5-HTT was successfully isolated and sequenced.
- The open reading frame encodes a 630-amino acid polypeptide with a molecular weight of approximately 70 kDa.
- Human 5-HTT shares 92% homology with rat 5-HTT but possesses a unique cAMP-dependent protein kinase phosphorylation site and lacks a conserved protein kinase C site.
Conclusions:
- The characterization of human 5-HTT provides insights into its molecular structure and potential regulation.
- Identified differences in phosphorylation sites suggest distinct regulatory mechanisms between human and rat serotonin transporters.