Random mutagenesis and screening of complex glycoproteins: expression of human gonadotropins in Dictyostelium discoideum.
The soil amoeba Dictyostelium discoideum is a host cell that provides simple genetics in combination with complex protein synthesis. We show that the complex human heterodimeric gonadotropins can be produced and secreted by this organism. Furthermore, both follicle stimulation hormone and choriogonadotropin produced by D. dictyostelium bind to their human receptors and elicit a biological response comparable to the wild-type hormones. We also show that structure-function analysis using random mutagenesis and screening of recombinant glycoprotein hormones is feasible. Thus, expression of gonadotropins in D. dictyostelium opens the way to the engineering of potential new therapeutic analogues.
Linskens
M H
MH
Cell Engineering Facility GBB, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands. m.linskens@chem.rug.nl
Grootenhuis
P D
PD
Blaauw
M
M
Huisman-de Winkel
B
B
Van Ravestein
A
A
Van Haastert
P J
PJ
Heikoop
J C
JC
D000818:Animals; D006063:Chorionic Gonadotropin / Q000096:biosynthesis / Q000235:genetics*; D004023:Dictyostelium / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005640:Follicle Stimulating Hormone / Q000096:biosynthesis / Q000235:genetics*; D005820:Genetic Testing; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D016296:Mutagenesis; D011994:Recombinant Proteins / Q000096:biosynthesis
D006063:Chorionic Gonadotropin; D006023:Glycoproteins; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone