Cole L A, Hartle R J, Laferla J J, Ruddon R W
Endocrinology. 1983 Sep;113(3):1176-8. doi: 10.1210/endo-113-3-1176.
Immunoaffinity adsorption techniques, utilizing specific antisera for hCG and its subunits bound to Sepharose 4B, have been employed to separate hCG alpha beta dimer and free subunits of hCG. As previously reported by this and a number of other laboratories, trophoblast cells (in vivo and in vitro) produce free alpha subunit in addition to hCG dimer. We have now shown that cultured JAr choriocarcinoma cells also secrete free beta subunit: 37% of the total beta subunit (combined and free) secreted by JAr cells is in the free form. Moreover, in pooled sera from choriocarcinoma patients 15% of total beta subunit is free, and in media from placental explant cultures and in pooled first trimester pregnancy sera 11% and 6.5%, respectively, of total beta subunit are in the free form. The free beta s are all of similar mol wt to the combined forms, and associate with urinary hCG alpha to form hCG. Free alpha s, which are larger than the combined forms, are unable to associate with urinary hCG beta to form hCG. We propose that the supply of combinable alpha subunit, rather than beta, limits dimer formation.
免疫亲和吸附技术利用与琼脂糖4B结合的针对人绒毛膜促性腺激素(hCG)及其亚基的特异性抗血清,已被用于分离hCGαβ二聚体和hCG的游离亚基。正如本实验室和其他一些实验室先前报道的那样,滋养层细胞(体内和体外)除了产生hCG二聚体外还产生游离的α亚基。我们现在已经表明,培养的JAr绒毛膜癌细胞也分泌游离的β亚基:JAr细胞分泌的总β亚基(结合型和游离型)中有37%是游离形式。此外,在绒毛膜癌患者的混合血清中,总β亚基的15%是游离的,在胎盘外植体培养物的培养基中以及在孕早期混合血清中,总β亚基分别有11%和6.5%是游离形式。游离的β亚基与结合形式的分子量相似,并与尿hCGα结合形成hCG。比结合形式更大的游离α亚基不能与尿hCGβ结合形成hCG。我们提出,可结合的α亚基而非β亚基的供应限制了二聚体的形成。