Effects of glycosylation on the conformation and dynamics of O-linked glycoproteins: carbon-13 NMR studies of ovine submaxillary mucin.
Carbon-13 NMR spectroscopic studies of native and sequentially deglycosylated ovine submaxillary mucin (OSM) have been performed to examine the effects of glycosylation on the conformation and dynamics of the peptide core of O-linked glycoproteins. OSM is a large nonglobular glycoprotein in which nearly one-third of the amino acid residues are Ser and Thr which are glycosylated by the alpha-Neu-NAc(2-6)alpha-GalNAc- disaccharide. The beta-carbon resonances of glycosylated Ser and Thr residues in intact and asialo mucin display considerable chemical shift heterogeneity which, upon the complete removal of carbohydrate, coalesces to single sharp resonances. This chemical shift heterogeneity is due to peptide sequence variability and is proposed to reflect the presence of sequence-dependent conformations of the peptide core. These different conformations are thought to be determined by steric interactions of the GalNAc residue with adjacent peptide residues. The absence of chemical shift heterogeneity in apo mucin is taken to indicate a loss in the peptide-carbohydrate steric interactions, consistent with a more relaxed random coiled structure. On the basis of the 13C relaxation behavior (T1 and NOE) the dynamics of the alpha-carbons appear to be unique to each amino acid type and glycosylation state, with alpha-carbon mobilities decreasing in the order Gly greater than Ala = Ser greater than Thr much greater than monoglycosylated Ser/Thr approximately greater than disaccharide linked Ser/Thr.(ABSTRACT TRUNCATED AT 250 WORDS)
Gerken
T A
TA
W. A. Bernbaum Center for Cystic Fibrosis Research, Cleveland, Ohio.
Butenhof
K J
KJ
Shogren
R
R
D000818:Animals; D002247:Carbon Isotopes; D006020:Glycopeptides; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009077:Mucins / Q000378:metabolism; D011487:Protein Conformation; D012756:Sheep; D013363:Submandibular Gland / Q000378:metabolism
D002247:Carbon Isotopes; D006020:Glycopeptides; D006023:Glycoproteins; D009077:Mucins