Functional characterization of the basolateral rat liver organic anion transporting polypeptide

Gerd Achim Kullak-Ublick, Bruno Hagenbuch, Bruno Stieger, Allan W. Wolkoff, Peter J. Meier

Research output: Contribution to journalArticle

159 Citations (Scopus)

Abstract

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 μmol/L) and taurocholate (Michaelis-Menten constant, 50 μmol/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 μmol/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, α-ketoglutarate and reduced glutathione were not taken up by organic aniontransporting 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. A variety of substrates previously shown to inhibit uptake of sulfobromophthalein and bile acids in perfused rat livers, isolated hepatocytes and basolateral membrane vesicles had no cis-inhibitory effects on organic anion-transporting polypeptide-mediated organic anion transport in X. laevis oocytes. Thus additional Na+-independent organic anion carriers must be present in the basolateral membranes of rat hepatocytes.

Original languageEnglish (US)
Pages (from-to)411-416
Number of pages6
JournalHepatology
Volume20
Issue number2
StatePublished - Aug 1994

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Anions
Sulfobromophthalein
Peptides
Liver
Indocyanine Green
p-Aminohippuric Acid
Oocytes
Hepatocytes
Complementary RNA
Taurocholic Acid
Xenopus laevis
Bile Acids and Salts
Glutathione
Taurochenodeoxycholic Acid
Glycocholic Acid
Organic Anion Transporters
Digitoxin
Cholates
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Bumetanide

ASJC Scopus subject areas

  • Hepatology

Cite this

Kullak-Ublick, G. A., Hagenbuch, B., Stieger, B., Wolkoff, A. W., & Meier, P. J. (1994). Functional characterization of the basolateral rat liver organic anion transporting polypeptide. Hepatology, 20(2), 411-416.

Functional characterization of the basolateral rat liver organic anion transporting polypeptide. / Kullak-Ublick, Gerd Achim; Hagenbuch, Bruno; Stieger, Bruno; Wolkoff, Allan W.; Meier, Peter J.

In: Hepatology, Vol. 20, No. 2, 08.1994, p. 411-416.

Research output: Contribution to journalArticle

Kullak-Ublick, GA, Hagenbuch, B, Stieger, B, Wolkoff, AW & Meier, PJ 1994, 'Functional characterization of the basolateral rat liver organic anion transporting polypeptide', Hepatology, vol. 20, no. 2, pp. 411-416.
Kullak-Ublick GA, Hagenbuch B, Stieger B, Wolkoff AW, Meier PJ. Functional characterization of the basolateral rat liver organic anion transporting polypeptide. Hepatology. 1994 Aug;20(2):411-416.
Kullak-Ublick, Gerd Achim ; Hagenbuch, Bruno ; Stieger, Bruno ; Wolkoff, Allan W. ; Meier, Peter J. / Functional characterization of the basolateral rat liver organic anion transporting polypeptide. In: Hepatology. 1994 ; Vol. 20, No. 2. pp. 411-416.
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abstract = "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 μmol/L) and taurocholate (Michaelis-Menten constant, 50 μmol/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 μmol/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, α-ketoglutarate and reduced glutathione were not taken up by organic aniontransporting 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. A variety of substrates previously shown to inhibit uptake of sulfobromophthalein and bile acids in perfused rat livers, isolated hepatocytes and basolateral membrane vesicles had no cis-inhibitory effects on organic anion-transporting polypeptide-mediated organic anion transport in X. laevis oocytes. Thus additional Na+-independent organic anion carriers must be present in the basolateral membranes of rat hepatocytes.",
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