Studies on the structure and biological function of the carbohydrate moiety of the extracellular matrix component laminin
Laminin from the EHS murine tumor was isolated as a noncovalent complex with entactin by lectin affinity chromatography using the ..cap alpha..-D-galactosyl binding lectin Griffonia simplicifolia I (GS I). Entactin was removed from this complex by passage over Sephacryl S-1000 in the presence of SDS. Laminin was found to contain 25-30% carbohydrate by weight. Free oligosaccharides were derived from the asparagine-linked glycans of affinity-purified laminin by hydrazinolysis, followed by re-N-acetylation and reduction with NaB(/sup 3/H)/sub 4/. The oligosaccharides were fractionated by affinity chromatography on concanavalin A (Con A)-Sepharose. A single peak corresponding to 20% of the total radioactivity was adsorbed, and specifically eluted with a linear gradient of 0-30 mM methyl ..cap alpha..-D-glucopyranoside. No additional material was eluted by 100 mM methyl ..cap alpha..-D-mannopyranoside. Further fractionation of the Con A-reactive oligosaccharides on GS I-Sepharose demonstrated that 70% of these oligosaccharides possess at least one terminal nonreducing ..cap alpha..-D-galactopyranosyl unit.
- Research Organization:
- Michigan Univ., Ann Arbor (USA)
- OSTI ID:
- 6281413
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AGGLUTININS
ANIMALS
ANTIBODIES
BIOLOGICAL FUNCTIONS
CARBOHYDRATES
CHROMATOGRAPHY
CONCANAVALIN
FRACTIONATION
FUNCTIONS
GLUCOPROTEINS
GLYCOPROTEINS
HEMAGGLUTININS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LECTINS
MAMMALS
MICE
MOLECULAR STRUCTURE
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
PROTEINS
RODENTS
SACCHARIDES
SEPARATION PROCESSES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES