Shao K, Balasubramanian S V, Pope C M, Bahl O P
State University of New York at Buffalo, Department of Biological Sciences, 14260-1300, USA.
Mol Cell Endocrinol. 1998 Nov 25;146(1-2):39-48. doi: 10.1016/s0303-7207(98)00194-4.
Human choriogonadotropin is a heterodimeric glycoprotein hormone comprised of noncovalently associated alpha- and beta-subunits. Each of the subunits has two N-glycosyl chains. In our previous communication, we investigated the role of individual carbohydrate chains in the alpha-subunit on the signal transduction function and conformation of the hormone. This paper deals with the effect of individual or both N-glycosyl chains in the beta-subunit on the function and conformation of the monomer as well as of the heterodimer. Three mutants each of hCGbeta and hCG lacking N-glycosyl chains at beta13Asn, beta30Asn and beta13,30Asn were prepared by site-directed mutagenesis by replacing Thr residues in the recognition triplet sequence at beta15 and beta32 positions with Gln. All mutant heterodimers had receptor binding and cAMP and progesterone stimulating activities comparable to wild type hCG. While the loss of carbohydrate at beta13Asn or beta13,30Asn in the case of hCGbeta monomer resulted in a 4-6%, decrease in the ordered structure, the loss of the glycosyl chain at beta 30Asn did not alter the conformation as compared with the wild type hCGbeta. Similarly, all carbohydrate deficient hCG heterodimers had a decrease of 6-8%) in the ordered structure as compared with hCG. Thus, while the individual N-glycosyl chains did not affect the function of the hormone, they did have marked effect on its conformation but the conformational changes were localized and did not perturb the receptor binding and signal transduction sites.
