EDTA-INSOLUBLE, CALCIUM-BINDING PROTEOGLYCAN IN BOVINE BONE
EDTA-INSOLUBLE, CALCIUM-BINDING PROTEOGLYCAN IN BOVINE BONE
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DOI：
10.1007/bf00301609
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发表时间：
1995-05-01
影响因子：
4.2
通讯作者：
YAMAUCHI, M
中科院分区：
文献类型：
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作者：
HASHIMOTO, Y;LESTER, GE;YAMAUCHI, M
已从脱矿物质、EDTA不溶的骨基质中分离出钙离子可沉淀、胰蛋白酶产生的蛋白聚糖片段。用胰蛋白酶完全消化脱矿物质基质，向上清液中加入浓度递增的CaCl 2，并分析所得沉淀。随着钙浓度的增加，沉淀物的量逐渐增加，并被EDTA可逆地溶解。经分子筛和阴离子交换层析后，得到一个含蛋白多糖的峰。免疫化学分析表明，该峰含有4-硫酸软骨素，可能还含有硫酸角质素。氨基酸分析表明，这种蛋白多糖含有大量的天冬氨酸/天冬酰胺（Asx），丝氨酸（Ser），谷氨酸/谷氨酰胺（Glx），脯氨酸（Pro）和甘氨酸（Gly）;然而，它含有很少的亮氨酸（Leu），这表明它不是富含亮氨酸的小蛋白多糖家族的成员。此外，显着量的磷酸丝氨酸（P-Ser）和羟脯氨酸（Hyp）的水解产物的这一部分。在SDS-PAGE上获得单一条带（Mr 59 kDa），其用考马斯亮蓝R-250染色而不用考马斯亮蓝R-250染色。如果骨粉在脱矿物质之前被胰蛋白酶消化，则该含蛋白聚糖的部分不会被释放。总的来说，这些结果表明，蛋白聚糖发生在脱矿基质中，其用CaCl 2沉淀，并与矿物质和胶原基质密切相关。这样的分子可能促进骨中诱导矿化的结构网络。
A calcium ion precipitable, trypsin-generated proteoglycan fragment has been isolated from the demineralized, EDTA-insoluble matrices of bone. The demineralized matrix was completely digested with trypsin, increasing concentrations of CaCl2 were added to the supernatant, and the resulting precipitates were analyzed. The amount of precipitate gradually increased with higher concentrations of calcium and was reversibly solubilized by EDTA. After molecular sieve and anion exchange chromatography, a proteoglycan-containing peak was obtained. Immunochemical analysis showed that this peak contained chondroitin 4-sulfate and possibly keratan sulfate. Amino acid analysis showed that this proteoglycan contained high amounts of aspartic acid/asparagine (Asx), serine (Ser), glutamic acid/glutamine (Glx), proline (Pro), and glycine (Gly); however, it contained little leucine (Leu) which suggests that it is not a member of the leucine-rich small proteoglycan family. In addition, significant amounts of phosphoserine (P-Ser) and hydroxyproline (Hyp) were identified in hydrolysates of this fraction. A single band (Mr 59 kDa) was obtained on SDS-PAGE that stained with Stains-all but not with Coomassie Brilliant Blue R-250. If bone powder was trypsinized prior to demineralization, this proteoglycan-containing fraction was not liberated. Collectively, these results indicate that a proteoglycan occurs in the demineralized matrix that is precipitated with CaCl2 and is closely associated with both mineral and collagen matrices. Such a molecule might facilitate the structural network for the induction of mineralization in bone.