Preparative electrophoresis on agarose submerged gels of two aggregating proteoglycan monomers from articular cartilage.
Analytical electrophoresis on polyacrylamide-agarose gels of aggregating proteoglycan monomers from baboon articular cartilage produces two distinct bands, corresponding to two different aggregating monomer populations. A preparative electrophoresis procedure is described for isolating the two monomers. Proteoglycans were extracted from young baboon articular cartilage in 4 M guanidinium chloride containing proteolysis inhibitors and aggregated after hyaluronic acid addition. The aggregates were separated from non-aggregated proteoglycans by isopycnic centrifugation, followed by gel chromatography on Sepharose CL-2B. The monomers of the aggregates were obtained by isopycnic centrifugation under dissociative conditions. Two monomers were separated by preparative electrophoresis on 0.8 % agarose submerged gels. Approximately 60 % of the proteoglycans were recovered from the gel using a freeze-squeeze procedure. Aliquots of the separated monomers gave single bands when submitted to analytical polyacrylamide-agarose gel electrophoresis. Their migration and appearance were similar to that of the two bands present in the non separated preparation of monomers.
D000818:Animals; D002358:Cartilage, Articular / Q000032:analysis*; D002500:Centrifugation, Isopycnic; D002850:Chromatography, Gel; D004586:Electrophoresis; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006820:Hyaluronic Acid; D046911:Macromolecular Substances; D010215:Papio; D011509:Proteoglycans / Q000302:isolation & purification*
D046911:Macromolecular Substances; D011509:Proteoglycans; D006820:Hyaluronic Acid