Roser J F, Papkoff H, Murthy H M, Chang Y S, Chloupek R C, Potes J A
Biol Reprod. 1984 Jun;30(5):1253-62. doi: 10.1095/biolreprod30.5.1253.
Donkey gonadotropins (donkey luteinizing hormone, dLH; donkey follicle-stimulating hormone, dFSH) have been isolated in purified form from 191 donkey pituitaries using essentially the same procedures previously employed for the purification of equine gonadotropins. Chemically, dLH and dFSH were observed to be similar to equine LH (eLH) and FSH (eFSH) in fractionation behavior and glycoprotein nature. Two forms of the dFSH molecule were observed, as is the case for eFSH. Donkey LH had significantly less total carbohydrate (13.5%) and sialic acid (1.9%) than eLH (26.7% and 5.8%, respectively). Carbohydrate (17-21%) and sialic acid (2.4%) content of the two dFSH preparations closely resembled that of eFSH. A slightly higher tyrosine content in the donkey gonadotropins was noted in a comparison of amino acid compositions. Immunologically, in a heterologous FSH radioimmunoassay (RIA), dFSH preparations were equal to or twice as active as eFSH preparations. However, in homologous RIAs for equine chorionic gonadotropin (eCG), eFSH and eLH, both the dLH and dFSH preparations were considerably less active than the equine gonadotropins, and their inhibition curves were all nonparallel. Biologically, in the Steelman-Pohley assay both dFSH preparations were equipotent and as potent as eFSH (approximately 40 times NIH-FSH-S12). In the Sertoli cell assay for cAMP (FSH assay) and the Leydig cell assay for testosterone (LH assay), both dFSH and dLH were 2- or 6-fold more active than eFSH and eLH, respectively. In rat and equine testis FSH homologous radioreceptor assays, dFSH preparations were as active and up to 6-fold more active than eFSH. In contrast, dLH was 10-fold less active than eLH in the equine LH homologous radioreceptor assay. Unlike eLH, dLH was found to possess little intrinsic FSH activity or FSH inhibitory activity, and the small amount of FSH activity observed was most likely due to FSH contamination. Therefore, eLH behaves much like eCG (pregnant mare's serum gonadotropin, PMSG) which also possesses both LH and FSH activity. In contrast, dLH behaves more like donkey chorionic gonadotropin (dCG) which possesses only a low degree of FSH activity.
已使用与先前用于纯化马促性腺激素基本相同的程序,从191个驴垂体中以纯化形式分离出驴促性腺激素(驴促黄体激素,dLH;驴促卵泡激素,dFSH)。在化学性质上,观察到dLH和dFSH在分级分离行为和糖蛋白性质方面与马促黄体激素(eLH)和促卵泡激素(eFSH)相似。观察到dFSH分子有两种形式,eFSH也是如此。驴促黄体激素的总碳水化合物(13.5%)和唾液酸(1.9%)含量明显低于马促黄体激素(分别为26.7%和5.8%)。两种dFSH制剂的碳水化合物(17 - 21%)和唾液酸(2.4%)含量与eFSH非常相似。在氨基酸组成比较中,注意到驴促性腺激素中的酪氨酸含量略高。在免疫方面,在异源促卵泡激素放射免疫分析(RIA)中,dFSH制剂的活性与eFSH制剂相当或为其两倍。然而,在马绒毛膜促性腺激素(eCG)、eFSH和eLH的同源RIA中,dLH和dFSH制剂的活性均明显低于马促性腺激素,且它们的抑制曲线均不平行。在生物学活性方面,在斯蒂尔曼 - 波利测定法中,两种dFSH制剂效力相当,且与eFSH效力相同(约为NIH - FSH - S12的40倍)。在用于检测环磷酸腺苷的支持细胞测定法(促卵泡激素测定)和用于检测睾酮的睾丸间质细胞测定法(促黄体激素测定)中,dFSH和dLH的活性分别比eFSH和eLH高2倍或6倍。在大鼠和马睾丸促卵泡激素同源放射受体测定中,dFSH制剂的活性与eFSH相当,且比eFSH高6倍。相比之下,在马促黄体激素同源放射受体测定中,dLH的活性比eLH低10倍。与eLH不同,发现dLH几乎不具有内在促卵泡激素活性或促卵泡激素抑制活性,观察到的少量促卵泡激素活性很可能是由于促卵泡激素污染。因此,eLH的行为很像eCG(孕马血清促性腺激素,PMSG),后者也同时具有促黄体激素和促卵泡激素活性。相比之下,dLH的行为更像驴绒毛膜促性腺激素(dCG),其仅具有低程度的促卵泡激素活性。