Preparation of a series of sulfated tetrasaccharides from shark cartilage chondroitin sulfate D using testicular hyaluronidase and structure determination by 500 MHz 1H NMR spectroscopy.
Six tetrasaccharide fractions were isolated from shark cartilage chondroitin sulfate D by gel filtration chromatography followed by HPLC on an amine-bound silica column after exhaustive digestion with testicular hyaluronidase. Their structures were determined unambiguously by one- and two-dimensional 500 MHz 1H NMR spectroscopy in conjunction with HPLC analysis of chondroitinase AC-II digests of the tetrasaccharides. One fraction was found to contain two tetrasaccharide components. All of the seven tetrasaccharides shared the common core structure GlcA beta 1-3GalNAc beta 1-4GLcA beta 1-3GalNAc with various sulfation profiles. Four were disulfated comprising of two monosulfated disaccharide units GLcA beta 1-3GalNAc(4-sulfate) and/or GlcA beta 1-3GalNAc(6-sulfate), whereas the other three were hitherto unreported trisulfated tetrasaccharides containing a disulfated disaccharide unit GlcA(2-sulfate)beta 1-3GalNAc(6-sulfate) and a monosulfated disaccharide unit GlcA beta 1-3GalNac(4- or 6-sulfate). These sulfated tetrasaccharides were demonstrated to serve as appropriate acceptor substrates for serum alpha-N-acetylgalactosaminyltransferase, indicating their usefulness as authentic oligosaccharide substrates or probes for the glycobiology of sulfated glycosaminoglycans.
Sugahara
K
K
Department of Biochemistry, Kobe Pharmaceutical University, Japan.
Tanaka
Y
Y
Yamada
S
S
D000818:Animals; D002236:Carbohydrate Conformation; D002356:Cartilage / Q000737:chemistry*; D002809:Chondroitin Sulfates / Q000737:chemistry*; D006821:Hyaluronoglucosaminidase / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009844:Oligosaccharides / Q000302:isolation & purification*; D012754:Sharks; D013737:Testis / Q000201:enzymology*
D009844:Oligosaccharides; C065068:chondroitin sulfate D; D002809:Chondroitin Sulfates; D006821:Hyaluronoglucosaminidase