Isolation and characterization of type IX collagen-proteoglycan from the Swarm rat chondrosarcoma.
Type IX collagen was partially purified from the Swarm rat chondrosarcoma by a series of a conventional salting-out procedures. The preparation was further separated by anion exchange chromatography into an unbound and a bound fraction in an A230 ratio of about 5:1. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the bound fraction appeared as a broad band, whose molecular mass ranged from 250 to 270 kDa. Digestion with chondroitinase ABC reduced the apparent molecular mass of the bound fraction to about 250 kDa, a value comparable to the molecular mass of the unbound fraction. Tryptic peptide maps of the protein moieties of unbound and bound forms showed that their molecular structures were basically identical. A monoclonal antibody specific for LMW, one of the pepsin-resistant fragments of the rat sarcoma type IX, reacted with both the unbound and bound fractions. Together the results indicate that the unbound and bound fractions represent a type IX collagen devoid of the chondroitin sulfate chain and its proteoglycan form with covalently bound chondroitin sulfate, respectively. The extent of glycosaminoglycan attachment to type IX collagen molecules in rat chondrosarcoma (about 16%) is quite different from the extents described in chick embryo cartilage (about 80%), chick vitreous humour (100%) and bovine cartilage (less than 5%). Further studies on the neoplastic tissue will offer additional information regarding the biological basis and biological consequences of the glycosaminoglycan attachment to type IX collagen molecules.
Arai
M
M
Institute for Molecular Science of Medicine, Aichi Medical University, Japan.
Yada
T
T
Suzuki
S
S
Kimata
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002813:Chondrosarcoma / Q000737:chemistry*; D003094:Collagen / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006025:Glycosaminoglycans / Q000032:analysis; D010434:Pepsin A; D010446:Peptide Fragments / Q000276:immunology; D011509:Proteoglycans / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains; D012513:Sarcoma, Experimental / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D006025:Glycosaminoglycans; D010446:Peptide Fragments; D011509:Proteoglycans; D003094:Collagen; D010434:Pepsin A