Application of 2-aminopyridine fluorescence labeling to glycosaminoglycans.
A pyridylamination method was applied to glycosaminoglycans and the characteristics of the resulting pyridylamino glycosaminoglycans were examined. First, glycosaminoglycan chains, which uniformly possess a xylose residue at their reducing termini, were liberated from proteoglycan by successive digestion with protease and endo-beta-xylosidase. Then the glycosaminoglycan chains were coupled with 2-aminopyridine by reductive amination with sodium cyanoborohydride for 15 h according to the method of Hase, S. et al. [J. Biochem. 95, 197-203 (1984)]. The pyridylamination reaction caused neither depolymerization, de-N-acetylation, nor de-N- or de-O-sulfation. The pyridylamino glycosaminoglycan chains had an intact linkage region (GlcA-Gal-Gal-Xyl) between the carbohydrate chain and the peptide core of the proteoglycan. These pyridylamino glycosaminoglycans should be useful as substrates for endo-type glycosidases that act on glycosaminoglycan chains and as markers for studies of glycosaminoglycan metabolism.
Department of Biochemistry, Hirosaki University School of Medicine, Aomori.
A
A
Kon
K
K
Takagaki
H
H
Kawasaki
T
T
Nakamura
M
M
Endo
D000631:Aminopyridines; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002809:Chondroitin Sulfates / Q000737:chemistry; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans / Q000737:chemistry*; D008969:Molecular Sequence Data; D014995:Xylosidases
D000631:Aminopyridines; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates; D014995:Xylosidases; C056547:endo-beta-xylosidase; C032439:alpha-aminopyridine