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


CONCLUSIONS
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.
Splinter
Patrick-L
PL
Center for Basic Research in Digestive Diseases, Mayo Clinic, Rochester, MN 55905, USA.
Lazaridis
Konstantinos-N
KN
Dawson
Paul-A
PA
LaRusso
Nicholas-F
NF
D000595:Amino Acid Sequence; D000818:Animals; D027321:Anion Transport Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001692:Biological Transport / Q000235:genetics / Q000502:physiology; D016466:CHO Cells; D027682:Cation Transport Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013656:Taurocholic Acid / Q000493:pharmacokinetics; D014162:Transfection; D000099173:Natural Resistance-associated Macrophage Protein 1
D027321:Anion Transport Proteins; D027682:Cation Transport Proteins; D004247:DNA; D012333:RNA, Messenger; D013656:Taurocholic Acid; D000099173:Natural Resistance-associated Macrophage Protein 1