TY - JOUR
T1 - Selective depolymerisation of keratan sulfate
T2 - production of radiolabelled substrates for 6-O-sulfogalactose sulfatase and β-d-galactosidase
AU - Hopwood, John J.
AU - Elliott, Helen
N1 - Funding Information:
This work was supported by grants from the Research Trust of the Adelaide Children’s Hospital Inc. and the National Health and Medical Research Council of Australia.
PY - 1983/6/16
Y1 - 1983/6/16
N2 - Keratan sulfate (KS) was N-deacetylated with anhydrous hydrazine and then degraded with nitrous acid, and the products were reduced with NaBT4. Radiolabelled disaccharides constituted at least 76% of the total oligosaccharide fraction. Three major disaccharides were isolated and identified. Of the total disaccharide isolated from bovine intervertebral-disc and human costal-cartilage, 91 and 79%, respectively, was identified as a disulfated disaccharide, O-(β-d-galactopyranosyl 6-sulfate)-(1→4)-2,5-anhydro-d-[1-3H]mannitol 6-sulfate (Gal6S-anM6S). The disaccharide fraction isolated from bovine-cornea KS contained only 14% of Gal6S-anM6S. The yield of monosulfated disaccharide, identified as O-β-d-galactopyranosyl-(1→4)-2,5-anhydro-d-[1- 3H]mannitol 6-sulfate, was 9, 17, and 84% of the total KS-disaccharide fraction isolated from intervertebral disc, costal cartilage, and cornea, respectively. For each of the KS type studied, the yield of unsulfated disaccharide was 4% of the total disaccharide-fraction. The tetrasaccharides were fractionated, on the basis of their sulfate content, into at least four species by paper electrophoresis, and some tentative structures are proposed. Disaccharide and tetrasaccharide species were evaluated as substrates for β-d-galactosidase and 6-O-sulfogalactose sulfatase.
AB - Keratan sulfate (KS) was N-deacetylated with anhydrous hydrazine and then degraded with nitrous acid, and the products were reduced with NaBT4. Radiolabelled disaccharides constituted at least 76% of the total oligosaccharide fraction. Three major disaccharides were isolated and identified. Of the total disaccharide isolated from bovine intervertebral-disc and human costal-cartilage, 91 and 79%, respectively, was identified as a disulfated disaccharide, O-(β-d-galactopyranosyl 6-sulfate)-(1→4)-2,5-anhydro-d-[1-3H]mannitol 6-sulfate (Gal6S-anM6S). The disaccharide fraction isolated from bovine-cornea KS contained only 14% of Gal6S-anM6S. The yield of monosulfated disaccharide, identified as O-β-d-galactopyranosyl-(1→4)-2,5-anhydro-d-[1- 3H]mannitol 6-sulfate, was 9, 17, and 84% of the total KS-disaccharide fraction isolated from intervertebral disc, costal cartilage, and cornea, respectively. For each of the KS type studied, the yield of unsulfated disaccharide was 4% of the total disaccharide-fraction. The tetrasaccharides were fractionated, on the basis of their sulfate content, into at least four species by paper electrophoresis, and some tentative structures are proposed. Disaccharide and tetrasaccharide species were evaluated as substrates for β-d-galactosidase and 6-O-sulfogalactose sulfatase.
UR - http://www.scopus.com/inward/record.url?scp=0021106140&partnerID=8YFLogxK
U2 - 10.1016/0008-6215(83)88092-6
DO - 10.1016/0008-6215(83)88092-6
M3 - Article
C2 - 6224558
AN - SCOPUS:0021106140
SN - 0008-6215
VL - 117
SP - 263
EP - 274
JO - Carbohydrate Research
JF - Carbohydrate Research
IS - C
ER -