Characterization of a large chondroitin sulfate proteoglycan present in bovine collateral ligament.
Bovine collateral ligament synthesized a 35S-labeled large proteoglycan species which eluted with a Kav of approximately 0.27 on Sepharose CL-2B and contained only chondroitin sulfate chains with a molecular mass of approximately 32 kDa. Fluorography of the 35S-labeled core proteins derived from the large ligament proteoglycan revealed a broad range of molecular masses above approximately 200 kDa, which was of comparable size to the four major endogenous core protein bands derived from this proteoglycan detected with 5/6/3-B-3, an antibody directed against terminal unsaturated chondroitin-6-sulfate disaccharides. The core proteins derived from the large ligament proteoglycan exhibited immunoreactivity of 12/21/1-C-6, an antibody specific for a peptide epitope common to both the G1 and G2 domains of aggrecan. Four major core protein bands with molecular masses greater than approximately 200 kDa derived from the large ligament proteoglycan, were detected using the antibodies raised against versican from bovine aorta or human fibroblasts. Compared with aggrecan, the 35S-labeled large ligament proteoglycan was distributed over a broader range of buoyant densities in an associative caesium chloride density gradient. This polydispersity may be indicative of differences in the degree of glycosylation as well as heterogeneity in the size of the large ligament proteoglycan core proteins. The 35S-labeled large ligament proteoglycan also demonstrated the ability to form complexes with an aggrecan aggregate preparation, the majority of which could not be dissociated by the presence of HA10-50. These findings indicate that the large chondrotin sulfate proteoglycan synthesized by bovine collateral ligament may be a versican-like proteoglycan which exhibited the potential to form like protein-stabilized complexes.
Campbell
M A
MA
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia. Margaret.Campbell@med.monash.edu.au
Tester
A M
AM
Handley
C J
CJ
Checkley
G J
GJ
Chow
G L
GL
Cant
A E
AE
Winter
A D
AD
Cain
W E
WE
D053674:Aggrecans; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D017885:Collateral Ligaments / Q000737:chemistry*; D004187:Disaccharides / Q000276:immunology; D016326:Extracellular Matrix Proteins; D066298:In Vitro Techniques; D037181:Lectins, C-Type; D008297:Male; D008970:Molecular Weight; D009844:Oligosaccharides / Q000494:pharmacology; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism
D053674:Aggrecans; D011508:Chondroitin Sulfate Proteoglycans; D004187:Disaccharides; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D009844:Oligosaccharides; D011509:Proteoglycans