TY - JOUR
T1 - Cloning and expression of SLC10A4, a putative organic anion transport protein
AU - Splinter, Patrick L.
AU - Lazaridis, Konstantinos N.
AU - Dawson, Paul A.
AU - LaRusso, Nicholas F.
PY - 2006/11/14
Y1 - 2006/11/14
N2 - 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.
AB - 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.
KW - Bile acid transport
KW - Biliary epithelium
KW - Plasma membrane
KW - SLC10A4
KW - Taurocholate
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U2 - 10.3748/wjg.v12.i42.6797
DO - 10.3748/wjg.v12.i42.6797
M3 - Article
C2 - 17106928
AN - SCOPUS:33751384466
SN - 1007-9327
VL - 12
SP - 6797
EP - 6805
JO - World journal of gastroenterology
JF - World journal of gastroenterology
IS - 42
ER -