Design of a long-acting follitropin agonist by fusing the C-terminal sequence of the chorionic gonadotropin beta subunit to the follitropin beta subunit.
Follitropin (FSH) is a pituitary glycoprotein hormone that is essential for the development of ovarian follicles and testicular seminiferous tubules. FSH is used clinically to stimulate follicular maturation for in vitro fertilization and treatment of anovulatory women. One issue regarding the clinical use of FSH is its short half-life in the circulation. To address this point, we constructed chimeric genes containing the sequence encoding the C-terminal peptide of the chorionic gonadotropin beta subunit (CG beta) fused to the translated sequence of the human FSH beta subunit (FSH beta). This region of CG beta is important for maintaining the prolonged plasma half-life of human CG dimer. The presence of the C-terminal peptide sequence did not significantly affect assembly of FSH beta with the alpha subunit or secretion of the dimer. In vitro receptor binding and steroidogenic activity of dimer bearing the FSH beta-C-terminal peptide chimera were the same as wild-type FSH. However, both the in vivo potency and half-life in circulation of the dimer bearing either one or two C-terminal peptide units were enhanced. Dimers containing FSH beta-CG beta chimeras could serve as potent FSH agonists for clinical use, and the present strategy may have wide applications for enhancing the in vivo half-life of diverse proteins.
Fares
F A
FA
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, MO 63110.
Suganuma
N
N
Nishimori
K
K
LaPolt
P S
PS
Hsueh
A J
AJ
Boime
I
I
D000818:Animals; D001141:Aromatase / Q000378:metabolism; D001681:Biological Assay; D016466:CHO Cells; D002478:Cells, Cultured; D002678:Chimera; D006063:Chorionic Gonadotropin / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D018997:Chorionic Gonadotropin, beta Subunit, Human; D006224:Cricetinae; D015195:Drug Design; D005260:Female; D005640:Follicle Stimulating Hormone / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism; D006207:Half-Life; D008297:Male; D008657:Metabolic Clearance Rate; D009929:Organ Size / Q000187:drug effects; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods*; D051381:Rats; D011962:Receptors, FSH / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism* / Q000494:pharmacology; D015183:Restriction Mapping; D013737:Testis / Q000378:metabolism; D014162:Transfection
D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D010446:Peptide Fragments; D011962:Receptors, FSH; D011993:Recombinant Fusion Proteins; D005640:Follicle Stimulating Hormone; D001141:Aromatase