A single N-acetylgalactosamine residue at threonine 106 modifies the dynamics and structure of interferon α2a around the glycosylation site

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DOIResolve DOI: http://doi.org/10.1074/jbc.M112.413252
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TypeArticle
Journal titleJournal of Biological Chemistry
ISSN0021-9258
Volume288
Issue1
Pages247254; # of pages: 8
SubjectEnhancement measurements; Heteronuclear; High resolution; Isotopically labeled; O-linked glycoproteins; Structure and dynamics; Three-dimensional structure; Time-scales; Amino acids; Dynamics; Escherichia coli; Esterification; Glucose; Glycosylation; Nuclear magnetic resonance spectroscopy; Glycoproteins; alpha2a interferon; glycoprotein; monosaccharide; n acetylgalactosamine; n acetylgalactosamine[13c,15n]alpha2a interferon; threonine; unclassified drug; addition reaction; article; biological activity; human; in vitro study; isotope labeling; molecular dynamics; nonhuman; nuclear magnetic resonance spectroscopy; priority journal; protein analysis; protein glycosylation; protein metabolism; protein polymorphism; protein structure; protein synthesis; Acetylgalactosamine; Computational Biology; Disulfides; Escherichia coli; Glycoproteins; Glycosylation; Humans; Interferon-alpha; Interferons; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Conformation; Peptides; Polysaccharides; Protein Conformation; Recombinant Proteins; Threonine; Escherichia coli
AbstractEnzymatic addition of GalNAc to isotopically labeled IFNα2a produced in Escherichia coli yielded the O-linked glycoprotein GalNAcα-[ 13C,15N]IFNα2a. The three-dimensional structure of GalNAcα-IFNα2a has been determined in solution by NMR spectroscopy at high resolution. Proton-nitrogen heteronuclear Overhauser enhancement measurements revealed that the addition of a single monosaccharide unit at Thr-106 significantly slowed motions of the glycosylation loop on the nanosecond time scale. Subsequent addition of a Gal unit produced Gal(β1,3) GalNAcα[13C,15N]IFNα2a. This extension resulted in a further decrease in the dynamics of this loop. The methodology used here allowed the first such description of the structure and dynamics of an O-glycoprotein and opens the way to the study of this class of proteins. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Biological Sciences (IBS-ISB)
Peer reviewedYes
NPARC number21269591
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Record identifiera8489825-4f72-4abc-8993-a85c2b2984fa
Record created2013-12-13
Record modified2016-05-09
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