Glycoconjugate Journal

, Volume 13, Issue 4, pp 609–619 | Cite as

Preparation of a series of sulfated tetrasaccharides from shark cartilage chondroitin sulfate D using testicular hyaluronidase and structure determination by 500 MHz1H NMR spectroscopy

  • Kazuyuki Sugahara
  • Yukako Tanaka
  • Shuhei Yamada
Glycoconjugate Journal

Abstract

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 MHz1H 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 the seven tetrasaccharides shared the common core structure GlcAβ1-3GalNAcβ1-4GlcAβ1-3GalNAc with various sulfation profiles. Four were disulfated comprising of two monosulfated disaccharide units GlcAβ1-3GalNAc(4-sulfate) and/or GlcAβ1-3GalNAc(6-sulfate), whereas the other three were hitherto unreported trisulfated tetrasaccharides containing a disulfated disaccharide unit GlcA(2-sulfate)β1-3GalNAc(6-sulfate) and a monosulfated disaccharide unit GlcAβ1-3GalNAc(4-or 6-sulfate). These sulfated tetrasaccharides were demonstrated to serve as appropriate acceptor substrates for serum α-N-acetylgalactosaminyltransferase, indicating their usefulness as authentic oligosaccharide substrates or probes for the glycobiology of sulfated glycosaminoglycans.

Keywords

chondroitin sulfate sulfated tetrasaccharides 500 MHz1H NMR 

Abbreviations

NFU

National formulary unit

COSY

correlation spectroscopy

HOHAHA

homonuclear Hartmann-Hahn

1D or 2D

one- or two-dimensional

IdoA

l-iduronic acid

ΔGlcA

d-gluco-4-enepyranosyluronic acid

ΔDi-0S

ΔGlcAβ1-3GalNAc

Di-4S

GlcAβ1-3GalNAc(4-sulfate)

ΔDi-4S

ΔGlcAβ1-3GalNAc(4-sulfate)

Di-6S

GlcAβ1-3GalNAc(6-sulfate)

ΔDi-6S

ΔGlcAβ1-3GalNAc(6-sulfate)

ΔDi-diSd

ΔGlcAβ(2-sulfate)α1-3GalNAc(6-sulfate)

ΔDi-diSE

ΔGlcAβ1-3GalNAc(4, 6-disulfate)

ΔU

G, U, 2S, 4S, and 6S represent ΔGlcA, GalNAc, GlcA, 2-O-sulfate, 4-O-sulfate, and 6-O-sulfate, respectively

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Copyright information

© Chapman & Hall 1996

Authors and Affiliations

  • Kazuyuki Sugahara
    • 1
  • Yukako Tanaka
    • 1
  • Shuhei Yamada
    • 1
  1. 1.Department of BiochemistryKobe Pharmaceutical UniversityKobeJapan

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