Purification and characterization of the human ovarian LH/hCG receptor and comparison of the properties of mammalian LH/hCG receptors.
Purification and characterization of the human ovarian LH/hCG receptor and comparison of the properties of mammalian LH/hCG receptors.
复制标题
人卵巢 LH/hCG 受体的纯化和表征以及哺乳动物 LH/hCG 受体特性的比较。
DOI：
10.1016/0006-2952(90)90240-l
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发表时间：
1990
影响因子：
5.8
通讯作者：
Wimalasena,J
中科院分区：
文献类型：
--
作者：
Alpaugh,K;Indrapichate,K;Abel,JA;Rimerman,R;Wimalasena,J
Methods previously published by us [Wimalasena et al., J Biol Chem260: 10689–10697, 1985; Wimalasena et al., J Biol Chem261: 9416–9420, 1986] were utilized to solubilize the human corpus luteal leuteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor with 3-[(3-cholamide-propyl) dimethylammonio]-1-propanesulfonate (CHAPS) and to purify the receptor by two steps of hCG-Sepharose affinity chromatography. The specific binding capacity (SBC) of the purified human receptor was 7510 pmol/mg protein, and K a= 2.2× 10 9 M− 1 when iodo hCG was competed by hCG; the yield was 4–7% of starting activity. When hLH was used in competition with hCG, specific binding capacity was 7900 pmol/mg protein and K a 1.0× 10 9 M− 1. Silver staining and autoradiography demonstrated a single protein of M r 78,000 under reducing and M r 58–62× 10 3 under nonreducing conditions. Rat ovarian LH/hCG receptor was purified by similar methods and the K a of 3.5× 10 10 M− 1 for hCG was substantially different from the K a for hLH which was 2.1× 10 9 M− 1. M r of the rat protein was 78–82× 10 3 (reduced) and 58–62× 10 3 (nonreduced) when analyzed by silver staining and autoradiography. For the first time, human LH/hCG receptor has been purified to apparent homogen-eity, and its M r of 78,000 was essentially identical to the M r values of purified rat and porcine receptors.
