The carboxyl-terminal region is a determinant for the intracellular behavior of the chorionic gonadotropin beta subunit: effects on the processing of the Asn-linked oligosaccharides.
The placental hormone human CG (hCG) consists of two noncovalently linked alpha- and beta-subunits similar to the other glycoprotein hormones LH, FSH, and TSH. These heterodimers share a common alpha subunit but differ in their structurally distinct beta subunits. The CGbeta subunit is distinguished among the beta subunits by the presence of a C-terminal extension with four serine-linked oligosaccharides (carboxyl terminal peptide or CTP). In previous studies we observed that deleting this sequence decreased assembly of the truncated CGbeta subunit (CGbeta114) with the alpha-subunit and increased the heterogeneity of the secreted forms of the uncombined subunit synthesized in transfected Chinese hamster ovary (CHO) cells. The latter result was attributed to alterations in the processing of the two N-linked oligosaccharides. To examine at what step this heterogeneity occurs, the CGbeta and CGbeta114 genes were transfected into wild-type and mutant CHO cell lines that are defective in the late steps of the N-linked carbohydrate-processing pathway. We show here that removal of the CTP alters the processing of the core mannosyl unit of the subunit to complex forms at both glycosylation sites and that the oligosaccharides contain polylactosamine. Although it has been presumed that there is little intramolecular interaction between the CTP and the proximal domains of the subunit, our data suggest that the CTP sequence participates in the folding of the newly synthesized subunit, which is manifest by the posttranslational changes observed here.
Muyan
M
M
Department of Molecular Biology & Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Boime
I
I
D000606:Amino Sugars / Q000096:biosynthesis; D000818:Animals; D001216:Asparagine / Q000378:metabolism*; D016466:CHO Cells; D002236:Carbohydrate Conformation; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006224:Cricetinae; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011134:Polysaccharides / Q000096:biosynthesis; D011499:Protein Processing, Post-Translational / Q000235:genetics; D017384:Sequence Deletion; D014162:Transfection
D000606:Amino Sugars; D018997:Chorionic Gonadotropin, beta Subunit, Human; D009844:Oligosaccharides; D010446:Peptide Fragments; D011134:Polysaccharides; C066929:polylactosamine; D001216:Asparagine