Fractionation of whale cartilage chondroitin sulfate on sepharose CL-4B in the presence of high concentration of ammonium sulfate. Relationship between molecular size and unit-disaccharide composition of the polysaccharide

✍ Scribed by Akira Ogamo; Atsuko Metori; Toshiharu Yamada; Kinzo Nagasawa

• Publisher: Elsevier Science
• Year: 1990
• Tongue: English
• Weight: 389 KB
• Volume: 198
• Category: Article
• ISSN: 0008-6215
• DOI: 10.1016/0008-6215(90)84287-5

✦ Synopsis

We reported',* that Sepharose CL-4B gel can retain sulfated glycosaminoglycuronans in concentrated ammonium sulfate solution at low temperature, resulting in an unique separation of the molecular species by stepwise elution of decreasing concentration of ammonium sulfate. The fractionation of shark cartilage chondroitin sulfate by this procedure2 indicated that the chondroitin sulfate molecules having the larger molecular size contain generally more chondroitin 6sulfate than chondroitin 4-sulfate units. Reciprocally, chondroitin sulfate having a smaller molecular size contains more chondroitin 4-sulfate than chondroitin B-sulfate units3. We describe herein the fractionation of whale cartilage chondroitin sulfate on a column of Sepharose CL4B with a reverse linear gradient of 3.25-1.7~ ammonium sulfate in 1OmM HCl at 4". The results confirme the correlation between molecular size and unit-disaccharide composition of the polysaccharide. EXPERIMENTAL Materials. -Whale cartilage chondroitin sulfate (special grade, lot No. W9082), chondroitinase ABC from Proteus vulgaris (chondroitin sulfate lyase, EC 4.2.99.6), and 4'S'-unsaturated disaccharide standards [ADi, 2-acetamido-2deoxy-3-0-(4-deoxy-a-L-threo-hex-4-enopyranosyluronic acid)-D-galactose; ADi-4S, 2-acetamido-2-deoxy-3-O-(Cdeoxy-a-L-threo-hex-4-enopyranosyluronic acid)-D-galactose 4-sulfate; ADi-6S, 2-acetamido-2-deoxy-3-0-(4-deoxy-cY-L-th-hex-4enopyranosyluronic acid)-D-galactose 6-sulfate; and ADi-diS,, 2-acetamido-2deoay-3-0-(4-deoxy-cY-L-threo-hex-4-enopyranosyluronic acid 2-or 3-sulfate)-D-*Chief research member of Kanagawa Prefectural Public Health Laboratories.