Single nucleotide polymorphism of the human high affinity choline transporter alters transport rate

Takashi Okuda, Michiko Okamura, China Kaitsuka, Tatsuya Haga, David Gurwitz

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

High affinity choline uptake plays a critical role in the regulation of acetylcholine synthesis in cholinergic neurons. Recently, we succeeded in molecular cloning of the high affinity choline transporter (CHT1), which is specifically expressed in cholinergic neurons. Here we demonstrate the presence of functionally relevant, nonsynonymous single nucleotide polymorphism in the human CHT1 gene by comprehensive sequence analysis of the exons and the intron/exon boundaries including the transcription start site. The deduced amino acid change for the polymorphism is isoleucine to valine at amino acid 89 (I89V) located within the third transmembrane domain of the protein. The allele frequency of I89V was 6% for Ashkenazi Jews. Functional assessment of the I89V transporter in mammalian cell lines revealed a 40-50% decrease in Vmax for choline uptake rate compared with the wild type, whereas there was no alteration in the apparent affinities for choline, sodium, chloride, and the specific inhibitor hemicholinum-3. There also was no change in the specific hemicholinum-3 binding activity. The decreased choline uptake was not associated with the surface expression level of the protein as assessed by biotinylation assay. These results suggest an impaired substrate translocation in the I89V transporter. The Caenorhabditis elegans ortholog of CHT1 has a valine residue at the corresponding position and a single replacement from valine to isoleucine caused a decrease in the choline uptake rate by 40%, suggesting that this hydrophobic residue is generally critical in the choline transport rate in CHT1. This polymorphism in the allelic CHT1 gene may represent a predisposing factor for cholinergic dysfunction.

Original languageEnglish
Pages (from-to)45315-45322
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number47
DOIs
Publication statusPublished - 2002 Nov 22
Externally publishedYes

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Choline
Polymorphism
Single Nucleotide Polymorphism
Nucleotides
Valine
Cholinergic Agents
Cholinergic Neurons
Isoleucine
Neurons
Exons
Functional assessment
Genes
Biotinylation
Amino Acids
Jews
Cloning
Transcription Initiation Site
Caenorhabditis elegans
Molecular Cloning
Gene Frequency

ASJC Scopus subject areas

  • Biochemistry

Cite this

Single nucleotide polymorphism of the human high affinity choline transporter alters transport rate. / Okuda, Takashi; Okamura, Michiko; Kaitsuka, China; Haga, Tatsuya; Gurwitz, David.

In: Journal of Biological Chemistry, Vol. 277, No. 47, 22.11.2002, p. 45315-45322.

Research output: Contribution to journalArticle

Okuda, Takashi ; Okamura, Michiko ; Kaitsuka, China ; Haga, Tatsuya ; Gurwitz, David. / Single nucleotide polymorphism of the human high affinity choline transporter alters transport rate. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 47. pp. 45315-45322.
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