Structural and Conformational Analysis of Glycan Moieties in Situ on Isotopically 13C,15N-Enriched Recombinant Human Chorionic Gonadotropin

Abstract
The conformational properties in solution of the glycans on the α subunit of recombinant human chorionic gonadotropin are described, using high-resolution multinuclear NMR studies on uniformly 13C,15N-enriched recombinant glycoprotein expressed in CHO cells. The glycan important for full biological activity of hCG, namely, that at Asn 52, appears to extend into solution both in the isolated α subunit and in complex with the β subunit. The disposition of this glycan with respect to the protein backbone suggests that glycosylation maintains full biological activity of hCG either by interacting with a lectin-like region of the hCG receptor or by reducing the affinity of the hormone for the hCG receptor and preventing its down-regulation.

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