Zherebtsova M A, Kazbekov E N
St. Petersburg Institute of Nuclear Physics of the Academy of Sciences, Gatchina, Russia.
Biochim Biophys Acta. 1994 Jun 1;1192(1):117-24. doi: 10.1016/0005-2736(94)90151-1.
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.
研究了溴乙酰对氨基马尿酸作为有机阴离子转运系统亲和探针的抑制特性。结果表明，溴乙酰化对氨基马尿酸能够不可逆地抑制刷状缘膜囊泡对氨基马尿酸（PAH）的摄取。这种抑制作用取决于处理时间和亲和探针浓度。用1 mM溴乙酰对氨基马尿酸处理刷状缘膜1.5小时，可导致PAH转运100%不可逆抑制，但碱性磷酸酶、γ-谷氨酰转肽酶或麦芽糖酶的活性未观察到变化。用溴乙酰对氨基[³H]马尿酸对有机阴离子转运体进行亲和标记。通过SDS-聚丙烯酰胺凝胶电泳表明，该探针与分子量为28 kDa、63 kDa、98 kDa和>150 kDa的刷状缘膜蛋白共价结合。以SITS和丙磺舒作为有机阴离子转运抑制剂获得的数据表明，28 kDa、63 kDa、98 kDa的刷状缘膜蛋白可能对应于有机阴离子转运系统。
