Equine follicle-stimulating hormone: molecular cloning of beta subunit and biological role of the asparagine-linked oligosaccharide at asparagine(56) of alpha subunit.
Equine FSH (eFSH) and eCG are members of the glycoprotein hormone family. These proteins are heterodimeric, composed of noncovalently associated alpha and beta subunits. We have previously reported that recombinant eCG has potent LH- and FSH-like activities and that the oligosaccharide at Asn(56) of the alpha subunit plays an indispensable role in expressing LH- but not FSH-like activity. In the present study, we cloned eFSH beta subunit cDNA and expressed wild-type recombinant eFSH and a partially deglycosylated mutant FSH (eFSH alpha56/beta) to investigate the biological role of the oligosaccharide at Asn(56) in FSH activity. The wild-type eFSH and eCG stimulated estradiol production in a dose-dependent manner in the primary cultures of rat granulosa cells, indicating that these equine gonadotropins have FSH activity. Partially deglycosylated eCG (eCG alpha56/beta) also stimulated estradiol production, confirming that the FSH-like activity of eCG is resistant to the removal of the N-linked oligosaccharide. Partially deglycosylated eFSH (eFSH alpha56/beta), however, did not show any FSH activity, indicating that the oligosaccharide at Asn(56) was necessary for eFSH. Thus, FSH-like activities of two gonadotropins, eCG and eFSH, are evoked through the distinct molecular mechanisms regarding the biological role of oligosaccharide at Asn(56) of the alpha subunit.
Saneyoshi
T
T
Laboratory of Cellular Biochemistry, Animal Resource Sciences/Veterinary Medical Sciences, University of Tokyo, Tokyo 113-8657, Japan.
Min
K S
KS
Jing Ma
X
X
Nambo
Y
Y
Hiyama
T
T
Tanaka
S
S
Shiota
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine / Q000737:chemistry*; D001483:Base Sequence; D002236:Carbohydrate Conformation; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000737:chemistry / Q000494:pharmacology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000235:genetics* / Q000494:pharmacology; D037201:Follicle Stimulating Hormone, beta Subunit; D006031:Glycosylation; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism; D006736:Horses / Q000235:genetics*; D007986:Luteinizing Hormone / Q000494:pharmacology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000502:physiology*; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D013329:Structure-Activity Relationship
D006063:Chorionic Gonadotropin; D018076:DNA, Complementary; D037201:Follicle Stimulating Hormone, beta Subunit; D009844:Oligosaccharides; D011994:Recombinant Proteins; D004958:Estradiol; D001216:Asparagine; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone