Specificity studies of bacterial sulfatases by means of structurally defined sulfated oligosaccharides isolated from shark cartilage chondroitin sulfate D.
Chondro-4-sulfatase and chondro-6-sulfatase from Proteus vulgaris and delta-hexuronate-2-sulfatase from Flavobacterium heparinum are potentially useful tools for structural studies of chondroitin sulfate and dermatan sulfate. Their substrate specificities were investigated with various structurally defined, sulfated hexasaccharides isolated from chondroitin sulfate as described in the accompanying report [Sugahara, K., Nadanaka, S., Takeda, K. & Kojima, T. (1996) Eur. J. Biochem. 239, 871-880]. The results indicated that delta-hexuronate-2-sulfatase released an ester sulfate from the C2 position of the delta-hexuronate residue located at the non-reducing terminus, while chondro-6-sulfatase removed an ester sulfate from the C6 position of the GalNAc residue at the reducing end of the hexasaccharides. Chondro-4-sulfatase acted preferentially on an ester sulfate on the C4 position of the GalNAc residue at the reducing end under mild incubation conditions, but also released a sulfate group under harsh conditions from the C4 position of the GalNAc residue at the internal positions of the hexasaccharide chains, unless the GalNAc residue had another ester sulfate on its C6 position. The results demonstrated the usefulness of the sulfatases as tools for the structural characterization of chondroitin sulfate oligosaccharides.
Sugahara
K
K
Department of Biochemistry, Kobe Pharmaceutical University, Japan.
Kojima
T
T
D000818:Animals; D002240:Carbohydrate Sequence; D002356:Cartilage / Q000737:chemistry; D002803:Chondro-4-Sulfatase; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000378:metabolism*; D005417:Flavobacterium / Q000201:enzymology; D006090:Gram-Negative Bacteria / Q000201:enzymology*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D011514:Proteus vulgaris / Q000201:enzymology; D012754:Sharks; D013379:Substrate Specificity; D013429:Sulfatases / Q000378:metabolism*
D009844:Oligosaccharides; C065068:chondroitin sulfate D; D002809:Chondroitin Sulfates; C101336:4-deoxy-threo-hex-4-enepyranosyluronic acid-2-sulfatase; D013429:Sulfatases; C046341:chondro-6-sulfatase; D002803:Chondro-4-Sulfatase