Protein engineering of a novel constitutively active hormone-receptor complex.

The Journal of Biological Chemistry
C WuD Puett

Abstract

Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of an alpha and a beta subunit that stimulates intracellular levels of cAMP via a G protein-coupled receptor. Herein we report the engineering and characterization of a novel molecule in which the receptor and its heterodimeric ligand were covalently linked in a single polypeptide chain. The hormone-receptor complex was expressed in cells transfected with this construct, but the cells were unable to bind significant amounts of exogenous hCG. However, cleavage of the hormone with a site-specific protease rendered the receptor accessible to exogenously added hormone. Cells transfected with the hCG-receptor construct contained elevated basal levels of cAMP; moreover, addition of hormone had no significant effect. These results are consistent with a strong and stable interaction between the single-chain hormone and its covalently linked receptor that results in a constitutively active complex.

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Citations

Feb 13, 2001·Trends in Endocrinology and Metabolism : TEM·V Garcia-Campayo, I Boime
Jan 19, 2010·Proceedings of the National Academy of Sciences of the United States of America·Adolfo Rivero-MüllerIlpo Huhtaniemi
Nov 12, 2009·The Journal of Biological Chemistry·Chia Lin ChangSheau Yu Teddy Hsu
Feb 10, 2006·Annals of the New York Academy of Sciences·David PuettPrema Narayan
Oct 21, 2015·Frontiers in Endocrinology·Prema Narayan
Oct 19, 2006·Molecular and Cellular Endocrinology·Thomas P Meehan, Prema Narayan
Apr 15, 2000·Methods : a Companion to Methods in Enzymology·P NarayanD Puett

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