The Presence of Fucose, Mannose, and Glucosamine-containing Heteropolysaccharide in Collagen from the Sea Anemone Metridium dianthus

Pepsin-solubilized collagen was prepared from the bodywall connective tissue of the sea anemone Mefridium dianfhus.It was judged to be collagen on the basis of its precipitability with 10% NaCl solution, its segment long spacing pattern in the electron microscope, its amino acid analysis, its melting behavior, its optical rotation, and its behavior on gel filtration chromatography, ion exchange chromatography, and disc electrophoresis.Pepsin-solubilized Metridium collagen contained two components, an a! chain and a /? component, when examined by polyacrylamide gel disc electrophoresis.These components were separated by agarose 1.5 m gel filtration and further purified by ion exchange chromatography on carboxymethylcellulose.The highly purified Q( and /? chains each contained almost 1% by weight of the sugars fucose, mannose, and glucosamine, in the form of heteropolysaccharide.These sugars have not been previously reported to be present in highly purmed, soluble collagen chains.In a previous communication we presented evidence that fucase, mannose, and glucosamine, in the form of heteropolysaccharide, were covalently linked via an asparaginyl-glucosamine linkage to a hydroxyproline-containing peptide (Peptide C) obtained from a collagenase digest of pepsin-solubilized collagen from the sea anemone Jdetridium dianthus (2).