A novel approach for chemically deglycosylating O-linked glycoproteins. The deglycosylation of submaxillary and respiratory mucins

Journal Article · Mon Jan 27 23:00:00 EST 1992 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00118a002· OSTI ID:5618329

Gerken, T A; Gupta, R; Jentoft, N ^[1]

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1. Case Western Reserve Univ., Cleveland, OH (United States)

A new approach for removing O-glycosidically linked carbohydrate side chains from glycoproteins is described. Periodate oxidation of the C3 and C4 carbons in peptide-linked N-acetylgalactosamine (GalNAc) residues generates a dialdehyde product which, under mild alkaline conditions, undergoes a {beta}-elimination which releases carbohydrate and leaves an intact peptide cores. The pH and time dependence, and intermediates of the elimination, have been extensively followed by carbon-13 NMR spectroscopy and amino acid analysis using ovine submaxillary mucin (OSM) as the substrate. The deglycosylation of OSM is complete and provides apomucin in high yield with an amino acid composition identical to the starting material. {sup 13}C NMR spectroscopic studies and carbohydrates analysis of the deglycosylation intermediates of the human mucin indicate that certain sialic acid containing and N-acetylglucosamine-containing oligosaccharides have elevated resistance to TFMSA treatment at 0C. By the use of neuraminidase, repeated mild TFMSA treatments, and multiple oxidations and {beta}-eliminations, the human mucin can be nearly completely deglycosylated. It is expected that all mucins and most glycoproteins containing O-glycosidic linkages can be readily and nearly completely deglycosylated using this combined approach.

• OSTI ID: 5618329
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 31:3; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOCHEMISTRY
CARBOHYDRATES
CARBON 13
CARBON ISOTOPES
CHEMICAL REACTIONS
CHEMISTRY
EVEN-ODD NUCLEI
GLYCOPROTEINS
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MUCOPROTEINS
NUCLEAR MAGNETIC RESONANCE
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXIDATION
POLYSACCHARIDES
PROTEINS
RESONANCE
SACCHARIDES
STABLE ISOTOPES
SULFONIC ACIDS