Purification and chemical composition of a low molecular weight follicle-stimulating hormone binding inhibitor from porcine follicular fluid.
Purification and chemical composition of a low molecular weight follicle-stimulating hormone binding inhibitor from porcine follicular fluid.
复制标题
猪卵泡液中低分子量卵泡刺激激素结合抑制剂的纯化和化学成分。
DOI：
10.1095/biolreprod41.5.863
复制
发表时间：
1989
影响因子：
3.6
通讯作者：
ReichertJr,LE
中科院分区：
文献类型：
--
作者：
Sluss,PM;Schneyer,AL;Andersen,TT;ReichertJr,LE
猪卵泡液含有多种能够在体外抑制促卵泡激素（FSH）与受体结合的因子，包括体外FSH生物活性的激动剂和拮抗剂。 FSH 受体结合抑制活性 (FSH-BI) 通过使用放射性配体（125 碘化物-人 FSH）受体（小牛睾丸膜）进行测定来确定；使用体外生物测定（培养的未成熟大鼠支持细胞）来确定拮抗剂/激动剂活性。 FSH 拮抗剂活性源自低分子量 (<5000) FSH-BI，可溶于酸性丙酮，不溶于乙醚，可进行制备规模分离。通过阴离子交换和反相高效液相色谱实现额外的纯化。高度纯化、具有生物活性的 FSH-BI 含有氨基酸 Ser、Gly、Arg、Thr、Ala、Pro、Val 和 Lys；己糖（苯酚-硫酸-阳性反应）；和乙醇胺。因此，这种 FSH 拮抗剂似乎是一种复杂的糖肽 - 可能源自膜成分，正如乙醇胺和碳水化合物残基的存在所表明的那样。因此，猪卵泡液含有低分子量 FSH 拮抗剂，与先前鉴定的高分子量 FSH 激动剂一起，可以通过与 FSH 受体的相互作用来调节性腺对 FSH 的反应。
Porcine follicular fluid contains several factors capable of inhibiting the binding, in vitro, of follicle-stimulating hormone (FSH) to receptor, including an agonist and an antagonist of FSH biological activity in vitro. FSH receptor-binding inhibitory activity (FSH-BI) was determined with assays using radioligand (125iodide-human FSH) receptor (calf-testes membrane); in vitro biological assays (cultured immature rat Sertoli cells) were used to determine antagonist/agonist activity. FSH antagonist activity is due to a low (<5000) molecular weight FSH-BI that is soluble in acidic acetone and insoluble in diethyl ether allowing preparative scale isolation. Additional purification was achieved by anion-exchange and reverse-phase high-performance liquid chromatography. Highly purified, biologically active FSH-BI contained the amino acids Ser, Gly, Arg, Thr, Ala, Pro, Val, and Lys; hexoses (phenol-sulfuric acid-positive reaction); and ethanolamine. Thus, this FSH antagonist appears to be a complex glycopeptide - possibly derived from membrane components, as suggested by the presence of ethanolamine and carbohydrate residues. Porcine follicular fluid, therefore, contains a low molecular weight FSH antagonist that, along with the high molecular weight FSH agonist previously identified, may regulate gonadal responsiveness to FSH through interactions with the FSH receptor.