title: Deciphering structural elements of mucin glycoprotein recognition.
abstract: Mucin glycoproteins present a complex structural landscape arising from the multiplicity of glycosylation patterns afforded by their numerous serine and threonine glycosylation sites, often in clusters, and with variations in respective glycans. To explore the structural complexities in such glycoconjugates, we used NMR to systematically analyze the conformational effects of glycosylation density within a cluster of sites. This allows correlation with molecular recognition through analysis of interactions between these and other glycopeptides, with antibodies, lectins, and sera, using a glycopeptide microarray. Selective antibody interactions with discrete conformational elements, reflecting aspects of the peptide and disposition of GalNAc residues, are observed. Our results help bridge the gap between conformational properties and molecular recognition of these molecules, with implications for their physiological roles. Features of the native mucin motifs impact their relative immunogenicity and are accurately encoded in the antibody binding site, with the conformational integrity being preserved in isolated glycopeptides, as reflected in the antibody binding profile to array components.
authors.affiliation: Center for Magnetic Resonance Research, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Borgert
authors.forename: Jamie
authors.initials: J
authors.lastname: Heimburg-Molinaro
authors.forename: Xuezheng
authors.initials: X
authors.lastname: Song
authors.forename: Yi
authors.initials: Y
authors.lastname: Lasanajak
authors.forename: Tongzhong
authors.initials: T
authors.lastname: Ju
authors.forename: Mian
authors.initials: M
authors.lastname: Liu
authors.forename: Pamela
authors.initials: P
authors.lastname: Thompson
authors.forename: Govind
authors.initials: G
authors.lastname: Ragupathi
authors.forename: George
authors.initials: G
authors.lastname: Barany
authors.forename: David F
authors.initials: DF
authors.lastname: Smith
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
authors.forename: David
authors.initials: D
authors.lastname: Live
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006020:Glycopeptides / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry* / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D040081:Protein Array Analysis; D011487:Protein Conformation; D013912:Threonine / Q000737:chemistry* / Q000378:metabolism
chemical_list: D006020:Glycopeptides; D009077:Mucins; D013912:Threonine