Structural Studies of Octasaccharides Derived from the Low-Sulfated Repeating Disaccharide Region and Octasaccharide Serines Derived from the Protein Linkage Region of Porcine Intestinal Heparin
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- 作者：Shuhei Yamada ; Kaori Sakamoto ; Hiromi Tsuda ; Keiichi Yoshida ; Makiko Sugiura ; and Kazuyuki Sugahara
- 刊名：Biochemistry
- 出版年：1999
- 出版时间：January 12, 1999
- 年：1999
- 卷：38
- 期：2
- 页码：838 - 847
- 全文大小：183K
- 年卷期：v.38,no.2(January 12, 1999)
- ISSN：1520-4995
文摘
Four octasaccharide serines and three octasaccharides were isolated after heparinase treatmentof porcine intestinal heparin. Their structures were characterized by enzymatic digestion in conjunctionwith HPLC and 500 MHz 1H NMR spectroscopy. Three of the four octasaccharide serines were structurallyidentical with those isolated previously, whereas one has the unreported structure 
HexA(2-sulfate)
1-4GlcN(N-sulfate)1-4GlcA
1-4GlcNAc1-4GlcA1-3Gal1-3Gal1-4Xyl1-O-Ser (HexA, GlcN,IdceA, and GlcA represent 4-deoxy--L-threo-hex-4-enepyranosyluronic acid, D-glucosamine, L-iduronicacid, and D-glucuronic acid, respectively). The other three octasaccharides were isolated for the first timeas discrete structures and shared the common core hexasulfated sequence HexA(2-sulfate)1-4GlcN(N-sulfate)1-4IdceA1-4GlcNAc1-4GlcA1-4GlcN(N-sulfate)1-4IdceA(2-sulfate)1-4GlcN(N,6-disulfate) with one or two additional sulfate groups. The octasaccharides which were derived fromthe low-sulfated repeating disaccharide region of heparin contained the common trisaccharide sequence-4IdceA1-4GlcNAc1-4GlcA1- [Yamada, S., Yamane, Y., Tsuda, H., Yoshida, K., and Sugahara,K. (1998) J. Biol. Chem. 273, 1863-1871], suggesting the programmed biosynthesis of heparin. Theseoctasaccharides are the largest oligosaccharides isolated so far from the low-sulfated irregular region ofheparin. Since oligosaccharides larger than a pentasaccharide appear to potentially exhibit binding activitiestoward growth factors or other functional proteins, they will be useful for investigating the structuralrequirement for molecular interactions between heparin and/or heparan sulfate and biologically activeproteins. During the course of the present structural studies, we evaluated the NMR data accumulatedthus far on heparin oligosaccharides and found several interesting rules on chemical shifts of proton signalsaffected by the neighboring sugar residues and their sulfation, which will be in turn useful for determiningstructures of unknown heparin and/or heparan sulfate oligosaccharides based on the proton resonances.
HexA(2-sulfate)1-4GlcN(N-sulfate)1-4GlcA1-4GlcNAc1-4GlcA1-3Gal1-3Gal1-4Xyl1-O-Ser (HexA, GlcN,IdceA, and GlcA represent 4-deoxy--L-threo-hex-4-enepyranosyluronic acid, D-glucosamine, L-iduronicacid, and D-glucuronic acid, respectively). The other three octasaccharides were isolated for the first timeas discrete structures and shared the common core hexasulfated sequence HexA(2-sulfate)1-4GlcN(N-sulfate)1-4IdceA1-4GlcNAc1-4GlcA1-4GlcN(N-sulfate)1-4IdceA(2-sulfate)1-4GlcN(N,6-disulfate) with one or two additional sulfate groups. The octasaccharides which were derived fromthe low-sulfated repeating disaccharide region of heparin contained the common trisaccharide sequence-4IdceA1-4GlcNAc1-4GlcA1- [Yamada, S., Yamane, Y., Tsuda, H., Yoshida, K., and Sugahara,K. (1998) J. Biol. Chem. 273, 1863-1871], suggesting the programmed biosynthesis of heparin. Theseoctasaccharides are the largest oligosaccharides isolated so far from the low-sulfated irregular region ofheparin. Since oligosaccharides larger than a pentasaccharide appear to potentially exhibit binding activitiestoward growth factors or other functional proteins, they will be useful for investigating the structuralrequirement for molecular interactions between heparin and/or heparan sulfate and biologically activeproteins. During the course of the present structural studies, we evaluated the NMR data accumulatedthus far on heparin oligosaccharides and found several interesting rules on chemical shifts of proton signalsaffected by the neighboring sugar residues and their sulfation, which will be in turn useful for determiningstructures of unknown heparin and/or heparan sulfate oligosaccharides based on the proton resonances.