Eggen K H, Malmstrøm A, Kolset S O
Norwegian Food Research Institute, As.
Biochim Biophys Acta. 1994 Feb 16;1204(2):287-97. doi: 10.1016/0167-4838(94)90020-5.
Proteoglycans were extracted and isolated from adult bovine muscle tissue by dissociative extraction followed by density gradient centrifugation, gel chromatography and ion-exchange chromatography. Two proteoglycans were characterized; one of large molecular size (PG-L) and one of small molecular size (PG-S). The recovery of PG-L and PG-S was 33% and 67%, respectively. By cellulose acetate electrophoresis before and after treatment with chondroitinase AC and ABC both samples were shown to carry predominantly dermatan sulfate chains. The large proteoglycan was recognized with an antibody against a large dermatan sulfate proteoglycan from bovine sclera, whereas the small was recognized by an antibody against decorin from bovine sclera. Chondroitinase ABC treatment of PG-S followed by SDS-PAGE showed a core protein with a molecular weight of 45 kDa, which also reacted with the decorin antibody. Amino-acid analysis of both PG-L and PG-S revealed an amino-acid composition closely similar, although not identical, to the large dermatan sulfate proteoglycan from bovine sclera and decorin, respectively. Immunohistochemical analyses of muscle tissue sections showed that decorin and the large dermatan sulfate proteoglycan are present in the perimysium layers of muscle tissue, although although with a somewhat different pattern of distribution. Decorin was, in addition, found in the endomysium.
通过解离提取，随后进行密度梯度离心、凝胶色谱和离子交换色谱，从成年牛肌肉组织中提取并分离出蛋白聚糖。鉴定出两种蛋白聚糖；一种是大分子尺寸的（PG-L），另一种是小分子尺寸的（PG-S）。PG-L和PG-S的回收率分别为33%和67%。在用软骨素酶AC和ABC处理之前和之后，通过醋酸纤维素电泳显示，两个样品主要携带硫酸皮肤素链。大分子蛋白聚糖可被抗牛巩膜大分子硫酸皮肤素蛋白聚糖的抗体识别，而小分子蛋白聚糖可被抗牛巩膜饰胶蛋白的抗体识别。用软骨素酶ABC处理PG-S后进行SDS-PAGE分析，显示出一种分子量为45 kDa的核心蛋白，该蛋白也与饰胶蛋白抗体发生反应。对PG-L和PG-S的氨基酸分析表明，它们的氨基酸组成分别与牛巩膜大分子硫酸皮肤素蛋白聚糖和饰胶蛋白密切相似，但并不完全相同。肌肉组织切片的免疫组织化学分析表明，饰胶蛋白和大分子硫酸皮肤素蛋白聚糖存在于肌肉组织的肌束膜层中，尽管分布模式略有不同。此外，在肌内膜中也发现了饰胶蛋白。
