Characteristic hexasaccharide sequences in octasaccharides derived from shark cartilage chondroitin sulfate D with a neurite outgrowth promoting activity
Characteristic hexasaccharide sequences in octasaccharides derived from shark cartilage chondroitin sulfate D with a neurite outgrowth promoting activity
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DOI：
10.1074/jbc.273.6.3296
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发表时间：
1998-02-06
影响因子：
4.8
通讯作者：
Sugahara, K
中科院分区：
文献类型：
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作者：
Nadanaka, S;Clement, A;Sugahara, K
小鼠脑硫酸软骨素（CS）蛋白聚糖，DSD-1-PG，具有DSD-1表位并且具有神经突生长促进特性。鲨鱼软骨CS-C抑制DSD-1特异性单克隆抗体473 HD和在小鼠神经胶质细胞上表达的DSD-1-PG的CS链之间的相互作用（Faissner，A.，Clement，A.，Lochter，A.，Streit，A.，曼德尔角，Schachner，M.（1994）J. Cell Biol.126，783-799），另一方面，从商业鲨鱼软骨CS-D分离的几种六糖，其含有较高比例的特征性D单元（GlcUA（2-硫酸盐）β 1-3GalNAc（6-硫酸盐））与CS-C相比，具有由A二糖单元（GlcUA β 1-3GalNAc（4-硫酸酯））和D二糖单元组成的A-D四糖序列（Nadanaka，S.和Sugahara，K.（1997）Glycobiology 7，253-263）。本文对鲨鱼软骨CS-D的生物活性和结构进行了研究。CS-D抑制单克隆抗体473 HD与DSD-1-PG的相互作用，并促进胚胎18天海马神经元突起生长。用细菌软骨素酶ABC部分消化CS-D后，通过凝胶过滤色谱和阴离子交换高效液相色谱分离出8个八糖组分，以研究A-D四糖单元在聚合物序列中的频率和排列。通过酶促双链反应与500 MHz H-1 NMR光谱的组合进行的结构分析表明，分离的八糖具有共同的核心结构Delta HexA α 1-3GalNAc β 1-4（GlcUA β 1-3GalNAc）（3）在不同的羟基上以不同的组合具有四个、五个和六个硫酸酯，在结构中，Δ HexA和GlcUA分别代表4-脱氧-α-L-苏型-己-4-烯吡喃糖醛酸和葡萄糖醛酸。没有发现D-D四糖序列，并且在代表约5.0%（w/w）的起始多糖的五种八糖中，离散的D二糖单元仅被证明为A-D-A或A-D-C六糖序列中的A-D四糖单元（C表示二糖GlcUA β 1-3GalNAc（6-硫酸盐））。它仍然有待确定是否存在于鲨鱼软骨CS-D中的这种特征性的六糖序列作为一些蛋白质配体识别的功能域结构。
A mouse brain chondroitin sulfate (CS) proteoglycan, DSD-1-PG, bears the DSD-1 epitope and has neurite out-growth promoting properties. Shark cartilage CS-C inhibits the interactions between the DSD-1-specific monoclonal antibody 473HD and the CS chains of the DSD-1-PG, which is expressed on the mouse glial cells (Faissner, A., Clement, A., Lochter, A., Streit, A., Mandl, C., and Schachner, M. (1994) J. Cell Biol. 126, 783-799), On the other hand, several hexasaccharides isolated from commercial shark cartilage CS-D, which contains a higher proportion of characteristic D units (GlcUA(2-sulfate)beta 1-3GalNAc(6-sulfate)) as compared with CS-C, has the A-D tetrasaccharide sequence composed of an A disaccharide unit (GlcUA beta 1-3GalNAc(4-sulfate)) and a D disaccharide unit (Nadanaka, S. and Sugahara, K. (1997) Glycobiology 7, 253-263). In this study, the biological activities and the structure of shark cartilage CS-D were investigated. CS-D inhibited the interactions between monoclonal antibody 473HD and DSD-1-PG and also promoted neurite outgrowth of embryonic day 18 hippocampal neurons. Eight octasaccharide fractions were isolated from CS-D after partial digestion with bacterial chondroitinase ABC by means of gel filtration chromatography and anion-exchange high performance liquid chromotography to investigate the frequency and the arrangement of the A-D tetrasaccharide unit in the polymer sequence. Structural analysis performed by a combination of enzymatic digestions with 500-MHz H-1 NMR spectroscopy demonstrated that the isolated octasaccharides shared the common core structure Delta HexA alpha 1-3GalNAc beta 1-4(GlcUA beta 1-3GalNAc)(3) with four, five, and six sulfate esters at various hydroxyl groups in different combinations, In the structure, Delta HexA and GlcUA represent 4-deoxy-alpha-L-threo-hex-4-enepyranosyluronic acid and glucuronic acid, respectively. No D-D tetrasaccharide sequence was found, and discrete D disaccharide units were demonstrated exclusively as A-D tetrasaccharide units in either an A-D-A or an A-D-C hexasaccharide sequence in the five octasaccharides that represented about 5.0% (w/w) of the starting polysaccharides (C denotes the disaccharide GlcUA beta 1-3GalNAc(6-sulfate)). It remains to be determined whether such characteristic hexasaccharide sequences present in shark cartilage CS-D serve as functional domain structures recognized by some protein ligands.