Miccio M, Baldini G, Basso V, Gazzin B, Lunazzi G C, Tiribelli C, Sottocasa G L
Dipartimento di Biochimica, University of Trieste, Italy.
Biochim Biophys Acta. 1989 May 19;981(1):115-20. doi: 10.1016/0005-2736(89)90088-6.
Monoclonal antibodies raised against bilitranslocase, may display either inhibitory or enhancing activity on the electrogenic transport of sulfobromophthalein, evoked in rat liver plasma-membrane vesicles by the addition of valinomycin in the presence of K+. In both cases, the target protein is identified with a 37 kDa band in SDS-mercaptoethanol gel electrophoresis of solubilized membranes. The electrophoretically homogeneous protein isolated by ion-exchange chromatography, corresponds in all respects to the 37 kDa protein band of bilitranslocase, obtained in the past by different techniques. Using this protein as antigen, a polyclonal monospecific antibody preparation has been obtained. As expected, the antibody preparation inhibits the electrogenic movement of sulfobromophthalein in plasma membrane vesicles from rat liver. It is concluded that the 37 kDa protein of bilitranslocase is at least a necessary component of the transport system involved in the sulfobromophthalein movement in plasma membrane.
针对胆红素转运酶产生的单克隆抗体，在存在钾离子的情况下添加缬氨霉素诱发大鼠肝细胞膜囊泡中磺溴酞的电转运时，可能表现出抑制或增强活性。在这两种情况下，目标蛋白在溶解膜的SDS-巯基乙醇凝胶电泳中均与一条37 kDa的条带一致。通过离子交换色谱法分离得到的电泳纯蛋白，在各方面均与过去通过不同技术获得的胆红素转运酶的37 kDa蛋白条带相符。以该蛋白作为抗原，获得了一种多克隆单特异性抗体制剂。正如预期的那样，该抗体制剂抑制了大鼠肝细胞膜囊泡中磺溴酞的电转运。得出的结论是，胆红素转运酶的37 kDa蛋白至少是参与磺溴酞在细胞膜中转运的运输系统的必要组成部分。
