Purification and characterization of dermatan sulfate from the skin of the eel, Anguilla japonica.
Glycosaminoglycans were isolated from the eel skin (Anguilla japonica) by actinase and endonuclease digestions, followed by a beta-elimination reaction and DEAE-Sephacel chromatography. Dermatan sulfate was the major glycosaminoglycan in the eel skin with 88% of the total uronic acid. The content of the IdoA2Salpha1-->4GalNAc4S sequence in eel skin, which shows anticoagulant activity through binding to heparin cofactor II, was two times higher than that of dermatan sulfate from porcine skin. The anti-IIa activity of eel skin dermatan sulfate was determined to be 2.4 units/mg, whereas dermatan sulfate from porcine skin shows 23.2 units/mg. The average molecular weight of dermatan sulfate was determined by gel chromatography on a TSKgel G3000SWXL column as 14 kDa. Based on 1H NMR spectroscopy, the presence of 3-sulfated and/or 2,3-sulfated IdoA residues was suggested. The reason why highly sulfated dermatan sulfate does not show anticoagulant activity is discussed. In addition to dermatan sulfate, the eel skin contained a small amount of keratan sulfate, which was identified by keratanase treatment.
Sakai
Shinobu
S
Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, 263-8522, Chiba, Japan.
Kim
Wan Seok
WS
Lee
In Sun
IS
Kim
Yeong Shik
YS
Nakamura
Ayako
A
Toida
Toshihiko
T
Imanari
Toshio
T
D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification*; D003871:Dermatan Sulfate / Q000737:chemistry / Q000302:isolation & purification*; D004187:Disaccharides / Q000032:analysis; D004524:Eels; D006025:Glycosaminoglycans / Q000302:isolation & purification; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular; D011516:Prothrombin / Q000037:antagonists & inhibitors; D012867:Skin / Q000737:chemistry*
D000925:Anticoagulants; D004187:Disaccharides; D006025:Glycosaminoglycans; D003871:Dermatan Sulfate; D011516:Prothrombin; C025337:Factor IIa