Hormone-specific inhibitory influence of (alpha)-subunit Asn56 oligosaccharide on in vitro subunit association and follicle- stimulating hormone receptor binding of equine gonadotropins

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 2841% 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