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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]