title: Isolation and characterization of a low molecular weight chondroitin sulfate proteoglycan from rabbit skeletal muscle.
abstract: Proteoglycans may be implicated in the process of aggregation of acetylcholine receptors in the basal lamina of skeletal muscle and possibly in the mechanism of reinnervation at the neuromuscular junction. In order to further deduce the role of such proteoglycans, we have sought to isolate them and define their molecular structures. In this study, proteoglycans were extracted from rabbit skeletal muscle by using 4 M guanidine hydrochloride and were purified by sequential cesium chloride density gradient ultracentrifugation, DEAE-cellulose ion-exchange chromatography, and Sepharose CL-6B and CL-2B gel filtration under dissociative conditions. A chondroitin sulfate proteoglycan which constituted about 44% of the total hexuronic acid content of the muscle tissue was isolated. This proteoglycan was found to have an apparent molecular weight [by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)] of 95,000, consistent with its small hydrodynamic size (Kav = 0.8 on Sepharose CL-2B), and to consist of peptide and glycosaminoglycan in a weight ratio of 1.0/0.8. The average molecular weight of its core protein-oligosaccharide remnants is 50,000, as estimated by SDS-PAGE of the chondroitinase ABC digested proteoglycan. Alkaline NaB3H4 treatment of the intact proteoglycan released chondroitin sulfate chains with an average molecular weight of 21,000. Pronase digestion of the intact proteoglycan generated glycosaminoglycan-peptides with an average of two chondroitin sulfate chains per peptide. These two saccharide units account for the total glycosaminoglycans per molecule and appear to be closely spaced on the core protein.(ABSTRACT TRUNCATED AT 250 WORDS)
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002499:Centrifugation, Density Gradient; D011508:Chondroitin Sulfate Proteoglycans / Q000302:isolation & purification*; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D006020:Glycopeptides / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000302:isolation & purification; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis; D011509:Proteoglycans / Q000302:isolation & purification*; D011817:Rabbits
chemical_list: D000596:Amino Acids; D011508:Chondroitin Sulfate Proteoglycans; D006020:Glycopeptides; D006025:Glycosaminoglycans; D009844:Oligosaccharides; D011509:Proteoglycans