title: Affinity identification of organic anion transporters in brush-border membrane vesicles from rat kidney.
abstract: The inhibitory properties of bromoacetyl-p-aminohippuric acid as the affinity probe of the organic anion transport system were studied. Bromoacetylated p-aminohippurate was shown to be able to inhibit irreversibly the p-aminohippurate (PAH) uptake in brush-border membrane vesicles. The inhibition depends on both the time of treatment and the affinity probe concentration. The treatment of brush-border membrane with 1 mM bromoacetyl-p-aminohippurate for 1.5 h results in 100% irreversible inhibition of PAH transport but no changes were observed in the activity of alkaline phosphatase, gamma-glutamyltranspeptidase or maltase. The affinity labelling of the organic anion transporters was performed with bromoacetyl-p-amino[3H]hippuric acid. It was shown, by means of SDS-polyacrylamide gel electrophoresis, that the probe bound covalently to the brush-border membrane proteins with molecular masses of 28 kDa, 63 kDa, 98 kDa, and > 150 kDa. The data obtained with SITS and probenecid as the organic anion transport inhibitors indicate that brush-border membrane proteins of 28 kDa, 63 kDa, 98 kDa may correspond to the organic anion transport system.
authors.affiliation: St. Petersburg Institute of Nuclear Physics of the Academy of Sciences, Gatchina, Russia.
authors.lastname: Orlov YuN
authors.forename: M A
authors.initials: MA
authors.lastname: Zherebtsova
authors.forename: E N
authors.initials: EN
authors.lastname: Kazbekov
mesh_terms: D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D000345:Affinity Labels; D000618:Aminohippuric Acids; D000818:Animals; D007672:Kidney Cortex / Q000378:metabolism* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis*; D008871:Microvilli / Q000201:enzymology / Q000378:metabolism*; D051381:Rats; D010130:p-Aminohippuric Acid / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D000618:Aminohippuric Acids; D008565:Membrane Proteins; C087730:bromoacetyl-4-aminohippuric acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D010130:p-Aminohippuric Acid