Functional characterization of the basolateral rat liver organic anion transporting polypeptide.
To characterize the transport functions of a recently cloned basolateral organic anion transporting polypeptide of rat hepatocytes we performed further kinetic transport and substrate cis-inhibition studies in organic anion-transporting polypeptide-cRNA injected Xenopus laevis oocytes. The studies demonstrate saturable Na(+)-independent sulfobromophthalein (Michaelis-Menten constant, 1.5 mumol/L) and taurocholate (Michaelis-Menten constant, 50 mumol/L) uptake by organic anion-transporting polypeptide. Sulfobromophthalein uptake was inhibited by the following organic anions: 0.01 mmol/L bilirubin (43%), 0.1 mmol/L indocyanine green (81%), 0.1 mmol/L 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS; 52%) and 1 mmol/L probenecid (74%). Competitive inhibition was shown for indocyanine green (inhibition constant about 1.3 mumol/L). Sulfobromophthalein and taurocholate uptakes were also inhibited by cholate, chenodeoxycholate, deoxycholate and ursodeoxycholate, as well as their glycine and taurine conjugates. Organic anion-transporting polypeptide also mediated uptake of glycocholate, tauroursodeoxycholate and taurochenodeoxycholate. No cis-inhibition of sulfobromophthalein uptake was seen in the presence of ATP, para-aminohippuric acid, bumetanide, digitoxin, reduced glutathione, leukotriene C4, nicotinic acid, ouabain, oxalate, rifampicin, succinate or sulfate. Furthermore, radioactively labeled para-aminohippuric acid, alpha-ketoglutarate and reduced glutathione were not taken up by organic anion-transporting polypeptide in cRNA-injected frog oocytes. These data confirm that organic anion-transporting polypeptide represents a novel hepatocellular organic anion uptake system that can mediate Na(+)-independent transport of monovalent (e.g., bile acids) and divalent (e.g., sulfobromophthalein and indocyanine green) cholephilic organic anions.(ABSTRACT TRUNCATED AT 250 WORDS)
Kullak-Ublick
G A
GA
Department of Medicine, University Hospital, Zürich, Switzerland.
Hagenbuch
B
B
Stieger
B
B
Wolkoff
A W
AW
Meier
P J
PJ
D000818:Animals; D027321:Anion Transport Proteins; D001647:Bile Acids and Salts / Q000493:pharmacokinetics; D002352:Carrier Proteins / Q000502:physiology*; D005260:Female; D066298:In Vitro Techniques; D007208:Indocyanine Green / Q000493:pharmacokinetics; D017136:Ion Transport; D008099:Liver / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D051381:Rats; D013448:Sulfobromophthalein / Q000493:pharmacokinetics; D014982:Xenopus laevis
D027321:Anion Transport Proteins; D001647:Bile Acids and Salts; D002352:Carrier Proteins; D013448:Sulfobromophthalein; D007208:Indocyanine Green