Purification and partial characterization of inhibin from porcine follicular fluid.
Purification and partial characterization of inhibin from porcine follicular fluid.
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猪卵泡液中抑制素的纯化和部分表征。
DOI：
10.1016/0006-291x(85)91849-2
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发表时间：
1985
影响因子：
3.1
通讯作者：
Vale,W
中科院分区：
文献类型：
--
作者：
Rivier,J;Spiess,J;McClintock,R;Vaughan,J;Vale,W
从猪卵泡液中分离纯化了抑制素，它是一种来源于性腺的蛋白质，可抑制垂体前叶细胞基础分泌卵泡刺激素。使用几个RP-HPLC步骤和在变性条件下的凝胶过滤，我们获得了大约一万倍纯化的级分，其在SDS PAGE上显示一条带，并且在相同的实验中，在还原后显示两条带（MW约14 K和约18 K），这表明对于拟南芥素的分子量为32 K。Edman降解表明，A链的前6个残基为H-Ser-Thr-Ala-Pro-Leu-Pro-，B链的前3个残基为H-Gly-Leu-Glu-。在我们的体外垂体细胞培养试验中，EC 50约为0.3 ng/ml或10 pM。产生了残基1-6的抗体，其可以在体外测定中免疫中和纯化的白藜芦碱活性。
Inhibin, a protein of gonadal origin that suppresses the basal secretion of follicle stimulating hormone by anterior pituitary cells has been purified from porcine follicular fluid. Using several RP-HPLC steps and gel filtration under denaturing conditions, we obtained a fraction approximately ten thousand fold purified which showed one band on SDS PAGE and in the same experiment two bands after reduction (MW ca 14K and ca 18K) suggesting a molecular weight of 32K for inhibin. Edman degradation of isolated inhibin and carboxymethylated chain A indicated that the first 6 residues were H-Ser-Thr-Ala-Pro-Leu-Pro-; by subtraction, the first 3 residues of chain B could be deduced to be H-Gly-Leu-Glu-. EC 50 was ca 0.3 ng/ml or 10 pM in our in vitro pituitary cell culture assay. Antibodies to residues 1–6 were raised which could immunoneutralize purified inhibin activity in an in vitro assay.