STRUCTURE AND INTERACTIONS OF CARTILAGE PROTEOGLYCAN BINDING REGION AND LINK PROTEIN
STRUCTURE AND INTERACTIONS OF CARTILAGE PROTEOGLYCAN BINDING REGION AND LINK PROTEIN
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DOI：
10.1042/bj2280077
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发表时间：
1985-01-01
影响因子：
4.1
通讯作者：
HARDINGHAM, TE
中科院分区：
文献类型：
--
作者：
BONNET, F;DUNHAM, DG;HARDINGHAM, TE
以猪喉软骨蛋白多糖为原料，经硫酸软骨素酶ABC和胰蛋白酶消化，制备了结合区和连接蛋白。凝胶层析实验表明，纯化的结合区与透明质酸（HA）可逆地相互作用，这种结合也被证明是由天然连接蛋白稳定。胰蛋白酶制备的链接蛋白质在溶液中的自关联的性质，部分抑制低聚糖（HA 10 -16）和废除的自由氨基（赖氨酸残基）与2-甲基马来酸酐的修改。修饰的连接蛋白的MW（沉降平衡）为41，700。结合区的分析表明，它含有25%（wt/wt）的碳水化合物，主要是半乳糖，葡萄糖胺，甘露糖和半乳糖胺。它含有一些硫酸角质素，如用内切-β-淀粉酶消化的那样。D-半乳糖苷酶（角蛋白酶）去除28%半乳糖和25%葡糖胺，MW从66，500降至60，800。角蛋白酶消化后，与结合区特异性多克隆抗体的相互作用不受影响，但用硫酸角蛋白单克隆抗体进行放射免疫测定的反应降低了47%。制备结合区、连接蛋白和.** 之间的复合物图形 **。显示MW为133，500的单一组分（5.5S）。在这种复合物中，连接蛋白的抗原决定簇出现掩蔽，如先前发现的蛋白聚糖聚集体。分离的结合区域和连接蛋白保留的性质与蛋白聚糖聚集体的结构和组织中所涉及的。
Binding region and link protein were prepared from pig laryngeal cartilage proteoglycans after chondroitinase ABC and trypsin digestion. Experiments on gel chromatography showed the purified binding region to interact reversibly with hyaluronate (HA), and this binding was also shown to be stabilized by native link protein. The trypsin-prepared link protein showed properties of self-association in solution that were partially inhibited by oligosaccharides (HA10-16) and abolished by modification of free amino groups (lysine residues) with 2-methylmaleic anhydride. The MW (sedimenation equilibrium) of the modified link protein was 41,700. Analysis of binding region showed it to contain 25% (wt/wt) carbohydrate, mainly in galactose, glucosamine, mannose and galactosamine. It contained some keratan sulfate, as digestion with endo-.beta.-D-galactosidase (keratanase) removed 28% galactose and 25% glucosamine and the MW decreased from 66,500 to 60,800. After keratanase digestion the interaction with polyclonal antibodies specific for binding region was unaffected, but the response in a radioimmunoassay with a monoclonal antibody to keratan sulfate was decreased by 47%. Preparation of a complex between binding region, link protein and .**GRAPHIC**. showed a single component (5.5S) of MW 133,500. In this complex the antigenic determinants of link protein appeared masked, as previously found with proteoglycan aggregates. The isolated binding region and link protein retained properties comparable with those involved in the structure and organization of proteoglycan aggregates.