Novel 70-kDa chondroitin sulfate/dermatan sulfate hybrid chains with a unique heterogenous sulfation pattern from shark skin, which exhibit neuritogenic activity and binding activities for growth factors and neurotrophic factors
Novel 70-kDa chondroitin sulfate/dermatan sulfate hybrid chains with a unique heterogenous sulfation pattern from shark skin, which exhibit neuritogenic activity and binding activities for growth factors and neurotrophic factors
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DOI：
10.1074/jbc.m412074200
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发表时间：
2005-02-11
影响因子：
4.8
通讯作者：
Sugahara, K
中科院分区：
文献类型：
--
作者：
Nandini, CD;Itoh, N;Sugahara, K
硫酸软骨素（CS）和硫酸皮肤素（DS）蛋白聚糖的混合链在生长因子结合、神经突发生和脑发育中至关重要。在这里，我们从鲨鱼皮中分离出CS/DS杂合链，旨在开发治疗剂。用各种软骨素酶消化表明，含有G1 cUA和IdoUA的二糖都沿着具有70 kDa的异常大的平均分子量的多糖链分散。通过阴离子交换色谱法将CS/DS链分离成主要（80%）和次要（20%）级分。这两个馏分有相对较低的硫酸化程度（硫酸/二糖摩尔比= 1.17与0.87），显示出独特的功能相比，海洋CS和DS分离到的日期，其中大部分是过硫酸化。它们是高度异质性的，特征在于多种二糖，包括G1 cUA-Ga 1 NAc、G1 cUA-Ga 1 NAc（6S）、GlcUA-Ga 1 NAc（4S）、IdoUA-Ga 1 NAc（4S）、G1 cUA-Ga 1 NAc（4S，6S）、IdoUA-Ga 1 NAc（4S，6S）、G1 cUA（2S）-Ga 1 NAc（6S）和/或IdoUA（2S）-Ga 1 NAc（6S）、IdoUA（2S）-Ga 1 NAc（4S）和新的GlcUA（2S）-Ga 1 NAc（4S），其中2S，4S，和6S分别代表2-O-、4-O-和6-O-硫酸根。CS/DS链结合两个神经营养因子和各种生长因子在大脑中表达的高亲和力的主要部分，通过动力学分析，使用表面等离子体共振检测器进行评估，也促进了轴突和树突性质的神经突的生长。用软骨素酶ABC、AC-1或B消化可完全消除神经突生成活性，表明含G1 cUA和IdoUA部分的重要性。它还显示出抗肝素辅因子II活性，与猪皮DS所表现出的活性相当。因此，凭借其独特的结构和生物活性，DS将在治疗方面找到潜在的用途。
Chondroitin sulfate (CS) and dermatan sulfate (DS) hybrid chains of proteoglycans are critical in growth factor binding, neuritogenesis, and brain development. Here we isolated CS/DS hybrid chains from shark skin aiming to develop therapeutic agents. Digestion with various chondroitinases showed that both G1cUA- and IdoUA-containing disaccharides are scattered along the polysaccharide chains with an unusually large average molecular mass of 70 kDa. The CS/DS chains were separated into major (80%) and minor (20% fractions by anion-exchange chromatography. Both fractions had relatively low degrees of sulfation (sulfate/disaccharide molar ratio = 1.17 versus 0.87), showing a unique feature compared with the marine CS and DS isolated to date, most of which are oversulfated. They were highly heterogeneous and characterized by multiple disaccharides including G1cUA-Ga1NAc, G1cUA-Ga1NAc(6S), GlcUA-Ga1NAc(4S), IdoUA-Ga1NAc(4S), G1cUA-Ga1NAc(4S,6S), IdoUA-Ga1NAc(4S,6S), G1cUA(2S)-Ga1NAc(6S), and/or IdoUA(2S)-Ga1NAc(6S), IdoUA(2S)-Ga1NAc(4S) and novel GlcUA(2S)-Ga1NAc(4S), where 2S, 4S, and 6S represent 2-O-, 4-O- and 6-O-sulfate, respectively. The CS/DS chains bound two neurotrophic factors and various growth factors expressed in the brain with high affinity as evaluated for the major fraction by kinetic analysis using a surface plasmon resonance detector, and also promoted the outgrowth of neurites of both an axonic and a dendritic nature. The neuritogenic activity was abolished completely by digestion with chondroitinase ABC, AC-I, or B, suggesting the importance of both G1cUA- and IdoUA-containing moieties. It also showed anti-heparin cofactor II activity comparable to that exhibited by DS from porcine skin. Thus, by virtue of its unique structure and biological activities, DS will find a potential use in therapeutics.