Cloning and expression of SLC10A4, a putative organic anion transport protein

Patrick L. Splinter, Konstantinos N Lazaridis, Paul A. Dawson, Nicholas F La Russo

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Aim: To determine if novel bile acid transporters may be expressed in human tissues. Methods: SLC10A1 (NTCP) was used as a probe to search the NCBI database for homology to previously uncharacterized ESTs. The homology search identified an EST (termed SLC10A4) that shares sequence identity with SLC10A1 and SLC10A2 (ASBT). We performed Northern blot analysis and RT-PCR to determine the tissue distribution of SLC10A4. SLC10A4 was cloned in frame with an epitope tag and overexpressed in CHO cells to determine cellular localization and functional analysis of bile acid uptake. Results: Northern analysis revealed that SLC10A4 mRNA is ubiquitously expressed in human tissues with the highest levels of mRNA expression in brain, placenta, and liver. In SLC10A4-transfected CHO cells, immunoblotting analysis and immunofluorescence staining demonstrated a 49-kDa protein that is expressed at the plasma membrane and intracellular compartments. Functional analysis of SLC10A4 showed no significant taurocholate uptake in the presence of sodium when compared to untransfected CHO cells. Conclusion: To date, we have shown that this protein has no capacity to transport taurocholate relative to SLC10A1; however, given its ubiquitous tissue distribution, it may play a more active role in transporting other endogenous organic anions.

Original languageEnglish (US)
Pages (from-to)6797-6805
Number of pages9
JournalWorld Journal of Gastroenterology
Volume12
Issue number42
StatePublished - Nov 14 2006

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Organic Anion Transporters
CHO Cells
Organism Cloning
Taurocholic Acid
Tissue Distribution
Messenger RNA
Bile Acids and Salts
Immunoblotting
Northern Blotting
Placenta
Fluorescent Antibody Technique
Anions
Epitopes
Proteins
Sodium
Cell Membrane
Databases
Staining and Labeling
Polymerase Chain Reaction
Liver

Keywords

  • Bile acid transport
  • Biliary epithelium
  • Plasma membrane
  • SLC10A4
  • Taurocholate

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Cloning and expression of SLC10A4, a putative organic anion transport protein. / Splinter, Patrick L.; Lazaridis, Konstantinos N; Dawson, Paul A.; La Russo, Nicholas F.

In: World Journal of Gastroenterology, Vol. 12, No. 42, 14.11.2006, p. 6797-6805.

Research output: Contribution to journalArticle

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AU - Dawson, Paul A.

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