Mass Spectrometric Evidence of Heparin Disaccharides for the Catalytic Characterization of a Novel Endolytic Heparinase
Ya-Peng Chao；Shao-Xiang Xiong；Xiu-Lan Cheng；Shi-Jun Qian
electrospray ionization-mass spectrometry ； heparin disaccharides ； heparinase ； high performance liquid chromatography ； Sphingobacterium sp.
Acta Biochimica et Biophysica Sinica
|Volume or Term/Year and Month of Publication||
36卷12期（2004 / 12 / 01）
840 - 844
Heparinase from different sources can eliminate heparin or/and heparan sulfate into various low-molecular weight heparins with different characteristics. Porcine intestinal mucosa heparin was degraded into a series of oligosaccharides by a novel heparinase from the species Sphingobacterium. Disaccharide components from the digests were separated and purified by ultrafiltration and HPLC. Five major peaks appeared as three types according to their retention time. The mass spectrometry of peak Ⅰ mainly gave the non-sulfated disaccharide with the mass of 379 Da. Peak Ⅱ and Ⅲ were indicated as two major monosulfated disaccharides with molecular mass of 417 and 459 Da respectively. Moreover, the peak Ⅲ represented an N-acetyl disaccharide. Both peak Ⅳ and Ⅴ showed the same mass of 496 Da, hinting that they were disulfate-substituted disaccharides. No trisulfate-substituted disaccharides were detected in the mixture of the heparin digest though they were abundant in the heparin structure. The results revealed that the heparinase might specifically cut the sites with low sulfated domain in heparin.
基礎與應用科學 > 化學
生物農學 > 生物科學