The biological action of choriogonadotropin is not dependent on the complete native quaternary interactions between the subunits.
Human CG (hCG) is a member of the glycoprotein hormone family characterized by a heterodimeric structure consisting of a common alpha-subunit noncovalently bound to a hormone-specific beta-subunit. The two subunits are highly intertwined and only the heterodimer is functional, implying that the quaternary structure is critical for biological activity. To assess the dependence of the bioactivity of hCG on the heterodimeric interactions, alpha- and beta-subunits bearing mutations that prevent assembly were covalently linked to form a single chain hCG. Receptor binding and signal transduction of these analogs were tested and their structural integrity analyzed using a panel of monoclonal antibodies (mAbs). These included dimer-specific mAbs, which react with at least four different epitope sites on the hormone, and some that react only with the free beta-subunit. We showed that there was significant loss of quaternary and tertiary structure in several regions of the molecule. This was most pronounced in single chains that had one of the disulfide bonds of the cystine knot disrupted in either the alpha- or beta-subunit. Despite these structural changes, the in vitro receptor binding and signal transduction of the single chain analogs were comparable to those of the nonmutated single chain, demonstrating that not all of the quaternary configuration of the hormone is necessary for biological activity.
Jackson
A M
AM
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St Louis, Missouri 63110, USA.
Berger
P
P
Pixley
M
M
Klein
C
C
Hsueh
A J
AJ
Boime
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D016466:CHO Cells; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D019281:Dimerization; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007668:Kidney; D008958:Models, Molecular; D015394:Molecular Structure; D016296:Mutagenesis; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D011974:Receptors, LH / Q000378:metabolism; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D014162:Transfection
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D000939:Epitopes; D011974:Receptors, LH