Selective removal of heparan sulfate chains from proteoheparan sulfate with a commercial preparation of heparitinase.
Procedures employing the commercial preparation of heparitinase were developed for isolating a protein-enriched core molecule from proteoheparan sulfate by selective removal of the heparan sulfate chains. Treatment of proteoheparan sulfate with the enzyme preparation caused seriously extensive degradation owing to the presence of proteolytic activity in the enzyme preparation. This effect could be avoided by using a series of protease inhibitors which prevented proteolytic degradation with less significant effect on the heparitinase activity. Application of the procedures to a purified preparation from the Engelbreth-Holm-Swarm tumor yielded a single protein-enriched core fraction with a molecular weight of approximately 450,000, as ascertained by sodium dodceyl sulfate-gel electrophoresis.
Kato
M
M
Oike
Y
Y
Suzuki
S
S
Kimata
K
K
D000818:Animals; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D006025:Glycosaminoglycans / Q000032:analysis* / Q000302:isolation & purification*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000032:analysis* / Q000302:isolation & purification*; D006868:Hydrolysis; D009374:Neoplasms, Experimental / Q000032:analysis; D011133:Polysaccharide-Lyases; D011480:Protease Inhibitors; D011509:Proteoglycans / Q000032:analysis*
D011508:Chondroitin Sulfate Proteoglycans; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011480:Protease Inhibitors; D011509:Proteoglycans; C021349:heparitinsulfate lyase