Butney V Y, Gotschall R R, Butnev V Y, Baker V L, Moore W T, Bousfield G R
Department of Biological Sciences, Wichita State University, Kansas 67260-0026, USA.
Biol Reprod. 1998 Feb;58(2):458-69. doi: 10.1095/biolreprod58.2.458.
Hybrid hormones were created using combinations of equine (e) LH, eFSH, and eCG alpha- and beta-subunit preparations. The efficiency of eFSH beta association was highest with eLH alpha (64-72%) and was lowest with eCG alpha (37-50%). Selective removal of alphaAsn56 oligosaccharide increased heterodimerization efficiency by 9-20% for eLH alpha, by 21-28% for eFSH alpha, and by 28-41% for eCG alpha. Both alpha and beta subunits contributed significantly to FSH receptor-binding activities of the hybrids. Purified hybrid hormone preparations consisting of either eFSH beta or eLH beta combined with eLH alpha, eFSH alpha, or eCG alpha were prepared. Equine FSH beta hybrids were more active in the FSH radioreceptor assay than eLH beta hybrids; within each beta-subunit group the eLH alpha hybrids were the most active, followed by eFSH alpha hybrids, while the least active were eCG alpha hybrids. A truncated, des(121-149) eLH beta derivative (eLH beta t) combined with native alpha-subunit preparations exhibited the same effect of alpha-subunit type on FSH receptor binding. Hybrids combining the eLH beta t derivative with Asn56-deglycosylated (N56dg-)eLH alpha, N56dg-eFSH alpha, and N56dg-eCG alpha preparations possessed 2.2- to 4.3-fold increased FSH receptor-binding activities as compared with the same hybrid preparations possessing the Asn56 carbohydrate. Granulosa cell bioassay of purified native eFSH beta and eLH beta hybrid hormones indicated no significant effect of the alpha-subunit carbohydrate differences on progesterone production. The alpha-subunit Asn56 oligosaccharide exerts a hormone-specific inhibitory influence on in vitro subunit reassociation and FSH receptor binding related to the size of its Man(alpha1-6)Man antenna.
使用马（e）促黄体生成素（LH）、促卵泡素（FSH）和人绒毛膜促性腺激素（eCG）α和β亚基制剂的组合制备了杂交激素。eFSHβ与eLHα结合的效率最高（64 - 72%），与eCGα结合的效率最低（37 - 50%）。选择性去除αAsn56寡糖后，eLHα的异二聚化效率提高了9 - 20%，eFSHα提高了21 - 28%，eCGα提高了28 - 41%。α和β亚基对杂交体的促卵泡素（FSH）受体结合活性均有显著贡献。制备了由eFSHβ或eLHβ与eLHα、eFSHα或eCGα组合而成的纯化杂交激素制剂。在FSH放射受体测定中，马促卵泡素β杂交体比促黄体生成素β杂交体更具活性；在每个β亚基组中，eLHα杂交体活性最高，其次是eFSHα杂交体，而活性最低的是eCGα杂交体。一种截短的、缺失（121 - 149）的eLHβ衍生物（eLHβt）与天然α亚基制剂结合，在FSH受体结合方面表现出相同的α亚基类型效应。将eLHβt衍生物与Asn56去糖基化（N56dg -）的eLHα、N56dg - eFSHα和N56dg - eCGα制剂组合而成的杂交体，与具有Asn56碳水化合物的相同杂交体制剂相比，FSH受体结合活性提高了2.2至4.3倍。纯化的天然eFSHβ和eLHβ杂交激素的颗粒细胞生物测定表明，α亚基碳水化合物差异对孕酮产生没有显著影响。α亚基Asn56寡糖对体外亚基重新结合和与FSH受体结合相关的激素特异性抑制作用与其Man（α1 - 6）Man天线的大小有关。
