Radhakrishnamurthy B, Jeansonne N, Berenson G S
Biochim Biophys Acta. 1986 Jun 3;882(1):85-96. doi: 10.1016/0304-4165(86)90059-0.
A chondroitin sulfate-dermatan sulfate proteoglycan was isolated from bovine aorta intima by extraction of the tissue by 4 M guanidine hydrochloride. The proteoglycan was purified by CsCl isopycnic centrifugation followed by gel filtration and ion-exchange chromatography. The proteoglycan had 21.9% protein, 22.1% uronate, 21.4% hexosamine and 10.8% sulfate. Glycosaminoglycan chains obtained from the proteoglycan by beta-elimination were resolved by gel filtration into two fractions, one containing chondroitin 6-sulfate with an approximate molecular weight of 49 000 and the other containing chondroitin 4-sulfate and dermatan sulfate in a proportion of 2:1 with an approximate molecular weight of 37 000. Digestion of the proteoglycan by chondroitinase ABC or AC yielded a protein core with similar composition and behavior in gel filtration and SDS-polyacrylamide gel electrophoresis. An approximate molecular weight of 180 000 was estimated for the core protein. Dermatan sulfate chains with an approximate molecular weight of 10 000 were observed only in the digest of chondroitinase AC. Limited trypsin hydrolysis of the proteoglycan yielded three peptide fragments containing chondroitin 6-sulfate, chondroitin 4-sulfate and dermatan sulfate in varied proportions. A tentative structure for the proteoglycan was suggested.
通过用4M盐酸胍提取组织，从牛主动脉内膜中分离出一种硫酸软骨素-硫酸皮肤素蛋白聚糖。该蛋白聚糖经氯化铯等密度离心，随后进行凝胶过滤和离子交换色谱法纯化。该蛋白聚糖含有21.9%的蛋白质、22.1%的糖醛酸、21.4%的己糖胺和10.8%的硫酸盐。通过β-消除法从该蛋白聚糖获得的糖胺聚糖链经凝胶过滤分离为两个组分，一个组分含有分子量约为49000的硫酸软骨素6-硫酸盐，另一个组分含有比例为2:1、分子量约为37000的硫酸软骨素4-硫酸盐和硫酸皮肤素。用软骨素酶ABC或AC消化该蛋白聚糖产生了一个在凝胶过滤和SDS-聚丙烯酰胺凝胶电泳中具有相似组成和行为的蛋白核心。估计核心蛋白的分子量约为180000。仅在软骨素酶AC的消化产物中观察到分子量约为10000的硫酸皮肤素链。用胰蛋白酶对该蛋白聚糖进行有限水解产生了三个肽片段，它们含有不同比例的硫酸软骨素6-硫酸盐、硫酸软骨素4-硫酸盐和硫酸皮肤素。提出了该蛋白聚糖的初步结构。
