http://www.abbs.info      E-mail: [email protected]

ISSN 1672-9145                                                Acta Biochim Biophys Sin 2004, 36(12): 840–844                                                  CN 31-1940/Q

Mass Spectrometric Evidence of Heparin Disaccharides for the Catalytic Characterization of A Novel Endolytic Heparinase

Yapeng CHAO1,3, Shaoxiang XIONG2, Xiulan CHENG1, and Shijun QIAN1*

 

1Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China;

2Center of Mass Spectrometry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China;

3Graduate School of the Chinese Academy of Sciences, Beijing 100039, China

 

Abstract        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.

 

Key words         electrospray ionization-mass spectrometry; heparin disaccharides; heparinase; high performance liquid chromatography; Sphingobacterium sp.

 

 

-----------------

Received: September 9, 2004        Accepted: November 15, 2004

Abbreviations: HPLC, high performance liquid chromatography; ESI-MS, electrospray ion-mass spectrometry; HLGAGs, heparin like glycosaminoglycans

This work was partially supported by the grants from the Chinese Academy of Sciences (KSCX2-3-02-03) and the National Natural Science Foundation of China (30270330)

*Corresponding author: Tel, 86-10-62651598; Fax, 86-10-62651598; E-mail, [email protected]