Eisen C, Meyer C, Wehling M
Division of Clinical Pharmacology, Medizinische Klinik, Klinikum Innenstadt, University of Munich, Germany.
Cell Mol Biol (Noisy-le-grand). 1997 Mar;43(2):165-73.
A new derivative of progesterone was synthesized for photoaffinity labelling of specific binding sites in porcine liver microsomes. Using progesterone-3-(O-carboxymethyl)-oximino-(3-125I-iodo-4-azidosa licylamidobutylamine) as a photoactivatable radioligand, selective covalent labelling of proteins was detected in porcine liver microsomes at apparent molecular weights of 90-100 kDa and 60-65 kDa by SDS-PAGE and subsequent radioautography. These proteins showed different ligand specifity for various steroids tested. On blue native polyacrylamide-gels three selectively labelled proteins were found corresponding to apparent molecular weights of approximately 310 kDa, approximately 215 kDa and approximately 75 kDa, respectively. Using two-dimensional electrophoresis for the analysis of these protein complexes, the 215 kDa-band could be correlated to the 90-100 kDa-band, while the 75 kDa-band may correspond to the 60-65 kDa-band at one-dimensional SDS-PAGE, respectively. One or more of these proteins may be involved in the rapid progesterone-induced increase of intracellular Ca2+ described previously in cultured hepatic cells.
合成了一种孕酮的新衍生物，用于对猪肝微粒体中的特异性结合位点进行光亲和标记。使用孕酮-3-(O-羧甲基)-肟基-(3-¹²⁵I-碘-4-叠氮水杨酰胺基丁胺)作为可光活化的放射性配体，通过SDS-PAGE和随后的放射自显影，在猪肝微粒体中检测到表观分子量为90-100 kDa和60-65 kDa的蛋白质的选择性共价标记。这些蛋白质对所测试的各种类固醇表现出不同的配体特异性。在蓝色天然聚丙烯酰胺凝胶上发现了三种选择性标记的蛋白质，其表观分子量分别约为310 kDa、约215 kDa和约75 kDa。使用二维电泳分析这些蛋白质复合物，在一维SDS-PAGE中，215 kDa条带可能与90-100 kDa条带相关，而75 kDa条带可能分别对应于60-65 kDa条带。这些蛋白质中的一种或多种可能参与了先前在培养的肝细胞中描述的孕酮诱导的细胞内Ca²⁺的快速增加。
