Molecular and functional characterization of an organic anion transporting polypeptide cloned from human liver.
OBJECTIVE
Based on a recently cloned rat liver organic anion transporter, we attempted to clone the corresponding human liver organic anion transporting polypeptide.


METHODS
A human liver complementary DNA library was screened with a specific rat liver complementary DNA probe. The human liver transporter was cloned by homology with the rat protein and functionally characterized in Xenopus laevis oocytes.


RESULTS
The cloned human liver organic anion transporting polypeptide consists of 670 amino acids and shows a 67% amino acid identity with the corresponding rat liver protein. Injection of in vitro transcribed complementary RNA into frog oocytes resulted in the expression of sodium-independent uptake of [35S]bromosulfophthalein (Michaelis constant [Km], approximately 20 mumol/L), [3H]cholate (Km, approximately 93 mumol/L), [3H]taurocholate (Km, approximately 60 mumol/L), [14C]glycocholate, [3H]taurochenodeoxycholate, and [3H]tauroursodeoxycholate (Km, approximately 19 mumol/L). Northern blot analysis showed cross-reactivity with messenger RNA species from human liver, brain, lung, kidney, and testes. Polymerase chain reaction analysis of genomic DNA from a panel of human-rodent somatic cell hybrids mapped the cloned human organic anion transporter to chromosome 12.


CONCLUSIONS
These studies show that the cloned human liver organic anion transporter is closely related to, but probably not identical to, the previously cloned rat liver transporter. Furthermore, its additional localization in a variety of extrahepatic tissues suggests that it plays a fundamental role in overall transepithelial organic anion transport of the human body.
Kullak-Ublick
G A
GA
Department of Medicine, University Hospital, Zürich, Switzerland.
Hagenbuch
B
B
Stieger
B
B
Schteingart
C D
CD
Hofmann
A F
AF
Wolkoff
A W
AW
Meier
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D027321:Anion Transport Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification*; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D015342:DNA Probes; D018076:DNA, Complementary; D006000:Glycocholic Acid / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009865:Oocytes / Q000737:chemistry; D051381:Rats; D013448:Sulfobromophthalein / Q000378:metabolism; D013655:Taurochenodeoxycholic Acid / Q000378:metabolism; D013657:Taurodeoxycholic Acid / Q000378:metabolism; D014982:Xenopus laevis
D027321:Anion Transport Proteins; D002352:Carrier Proteins; D015342:DNA Probes; D018076:DNA, Complementary; D013448:Sulfobromophthalein; D013655:Taurochenodeoxycholic Acid; D013657:Taurodeoxycholic Acid; D006000:Glycocholic Acid