Isolation and Structural Determination of Novel Sulfated Hexasaccharides from Squid Cartilage Chondroitin Sulfate E That Exhibits Neuroregulatory Activities
详细信息 查看全文
- 作者：Akiko Kinoshita ; Shuhei Yamada ; Stuart M. Haslam ; Howard R. Morris ; Anne Dell ; and Kazuyuki Sugahara
- 刊名：Biochemistry
- 出版年：2001
- 出版时间：October 23, 2001
- 年：2001
- 卷：40
- 期：42
- 页码：12654 - 12665
- 全文大小：202K
- 年卷期：v.40,no.42(October 23, 2001)
- ISSN：1520-4995
文摘
Squid cartilage chondroitin sulfate E (CS-E) exhibits various biological activities, includinganticoagulant activities, lymphoid regulatory activities, and neuroregulatory activities [Ueoka, C., Kaneda,N., Okazaki, I., Nadanaka, S., Muramatsu, T., and Sugahara, K. (2000) J. Biol. Chem. 275, 37407-37413]. These activities are expressed through molecular interactions with specific proteins, includingheparin cofactor II, selectins, CD44, chemokines, and the heparin-binding growth factor midkine. Hence,the sugar sequence information is essential for a better understanding of the CS-E functions. Previously,several novel tetrasaccharides containing the unreported 3-O-sulfated glucuronic acid (GlcA) were isolatedafter digestion of squid cartilage CS-E with testicular hyaluronidase. In this study, hexasaccharides wereisolated to obtain more detailed sequence information, especially around the GlcA(3-O-sulfate) residue,and were characterized by fast atom bombardment mass spectrometry and 500 or 600 MHz 1H NMRspectroscopy. The findings demonstrate one tetrasulfated and five pentasulfated hexasaccharide sequences,five of them being novel. They were composed of three disaccharide building units of either A [GlcA(
1-3)GalNAc(4-O-sulfate)], E [GlcA(1-3)GalNAc(4,6-O-disulfate)], K [GlcA(3-O-sulfate)(1-3)GalNAc(4-O-sulfate)], L [GlcA(3-O-sulfate)(1-3)GalNAc(6-O-sulfate)], or M [GlcA(3-O-sulfate)(1-3)GalNAc(4,6-O-disulfate)], forming E-A-A, M-A-A, K-L-A, E-E-A, K-K-A, and A-M-A hexasaccharidesequences. The K-L tetrasaccharide sequence is to date unreported. The isolated sequences appear toindicate the occurrence of an unreported GlcA 3-O-sulfotransferase specific for chondroitin sulfate. Theobtained sequence information will be useful for investigating the structure-function relationship andbiosynthesis of CS-E.
1-3)GalNAc(4-O-sulfate)], E [GlcA(1-3)GalNAc(4,6-O-disulfate)], K [GlcA(3-O-sulfate)(1-3)GalNAc(4-O-sulfate)], L [GlcA(3-O-sulfate)(1-3)GalNAc(6-O-sulfate)], or M [GlcA(3-O-sulfate)(1-3)GalNAc(4,6-O-disulfate)], forming E-A-A, M-A-A, K-L-A, E-E-A, K-K-A, and A-M-A hexasaccharidesequences. The K-L tetrasaccharide sequence is to date unreported. The isolated sequences appear toindicate the occurrence of an unreported GlcA 3-O-sulfotransferase specific for chondroitin sulfate. Theobtained sequence information will be useful for investigating the structure-function relationship andbiosynthesis of CS-E.