Biosynthesis of a biologically active single peptide chain containing the human common alpha and chorionic gonadotropin beta subunits in tandem.
One of the distinguishing features of the gonadotropin and thyrotropin hormone family is their heterodimeric structure, consisting of a common alpha subunit and a hormone-specific beta subunit. Subunit assembly is vital to the function of these hormones: The conformation of the heterodimer is essential for controlling secretion, hormone-specific posttranslational modifications, and signal transduction. To address whether alpha and beta subunits can be synthesized as one chain and also maintain biological activity, a chimera composed of the human chorionic gonadotropin (hCG) beta subunit genetically fused to the alpha subunit was constructed. The resulting polypeptide hCG molecule not only was efficiently secreted but also displayed an increased biological activity in vitro and in vivo. These data show that the alpha and hCG beta subunits encoded as a single chain retain a biologically active conformation similar to that seen in the heterodimer. This approach can be used to investigate structure-function relationships of the glycoprotein hormone family that were previously not tractable because of the absolute dependence on assembly for the biological response. Moreover, other bioactive multisubunit ligands can be engineered where the combination efficiency and specificity of heterodimers and homodimers are otherwise difficult to control.
Sugahara
T
T
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, MO 63110.
Pixley
M R
MR
Minami
S
S
Perlas
E
E
Ben-Menahem
D
D
Hsueh
A J
AJ
Boime
I
I
D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D016466:CHO Cells; D006063:Chorionic Gonadotropin / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D006224:Cricetinae; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D005091:Exons; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016296:Mutagenesis; D006080:Ovarian Follicle / Q000187:drug effects / Q000502:physiology; D010060:Ovulation / Q000187:drug effects; D016133:Polymerase Chain Reaction; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D014162:Transfection
D006063:Chorionic Gonadotropin; D017931:DNA Primers; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone