Genetic engineering of single-chain gonadotropins and hormone-receptor fusion proteins.
The gonadotropin hormone family is distinguished by its heterodimeric structure in which the members share a common alpha subunit and a hormone-specific beta subunit. Since assembly of the heterodimer is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking the two subunits. The single-chain hormone can in turn be fused to its receptor to produce a functional single-chain hormone-receptor complex. These fusion constructs offer a valuable new approach in structure-function studies and in the generation of hormone analogs. In this article we describe the experimental design for the generation of single-chain human chorionic gonadotropin and single-chain hormone-receptor fusion complex and strategies for the expression of these fusion proteins.
Narayan
P
P
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA. narayan@bchiris.bmb.uga.edu
Wu
C
C
Puett
D
D
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000737:chemistry / Q000235:genetics; D000242:Cyclic AMP / Q000096:biosynthesis; D005818:Genetic Engineering / Q000379:methods*; D006062:Gonadotropins / Q000138:chemical synthesis* / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D011967:Receptors, Gonadotropin / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D006063:Chorionic Gonadotropin; D006062:Gonadotropins; D010455:Peptides; D011967:Receptors, Gonadotropin; D011993:Recombinant Fusion Proteins; D000242:Cyclic AMP