Affinity binding of the cartilage proteoglycan protein-keratan sulfate core to immobilized hyaluronic acid

✍ Scribed by James E. Christner; Martin L. Brown; Dominic D. Dziewiatkowski

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 679 KB
• Volume: 90
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90004-0

✦ Synopsis

The protein-keratan sulfate core of bovine nasal cartilage proteoglycan was purified by affinity chromatography on a column of immobilized hyaluronic acid. The hyaluronic acid was immobilized by reaction with a hydrazido-alkyl derivative of Sepharose in the presence of borohydride. Proteoglycan was digested with chondroitinase ABC and the entire mixture was passed over a column of the Sepharose-hyaluronic acid maintained at 4°C. After the digested chondroitin sulfate chains were washed from the column, the bound protein-keratan sulfate core was eluted with 4 M guanidinium chloride. The protein-keratan sulfate core interacts with the affinity matrix through its hyaluronic acid binding site as shown by the inhibition of binding by free hyaluronic acid and hyaluronic acid decasaccharide.

I Abbreviations used: HA, hyaluronic acid; Sepharose-HA, hyaluronic acid covalently attached to adipic acid dihydrazide-substituted Sepharose 2B; GuHCl, guanidinium hydrochloride; core, protein-keratan sulfate core prepared from chondroitinase ABD-digested proteoglycan.