Vogel K G, Heinegård D
J Biol Chem. 1985 Aug 5;260(16):9298-306.
Proteoglycans were extracted in good yield from the proximal, fibrous portion of adult bovine tendon with 4 m guanidine HCl. They comprise less than 1% of the dry weight of the tissue. Using CsCl density gradient centrifugation, gel chromatography, and ion exchange chromatography, two populations of proteoglycans were separated and purified from other tissue proteins. One was a large, chondroitin sulfate proteoglycan with high buoyant density in CsCl. This component appeared to be composed of two or three subpopulations as detected by agarose/polyacrylamide electrophoresis, although they could not be effectively separated from one another for individual characterization. As a group, the large proteoglycans eluted from Sepharose CL-2B with Kav from 0.1-0.5 and their core protein had Mr greater than 200,000 with high contents of glutamic acid, serine, and glycine. The glycosaminoglycan chains had a weight average Mr of 17,000 and more than 98% of the uronic acid was glucuronic acid. This group comprised only 12% of the total proteoglycan of the tissue. The other 88% of the proteoglycans appeared to represent one group of small molecules that eluted from Sepharose CL-2B at Kav = 0.70. They demonstrated buoyant densities in a CsCl gradient ranging from greater than or equal to 1.51 to 1.30 g/ml. Their core protein had an apparent Mr = 48,000 following removal of the glycosaminoglycan chains by digestion with chondroitinase ABC. This core protein had a particularly high content of aspartic acid/asparagine and leucine. The glycosaminoglycan chains had a weight average Mr of 37,000 and were dermatan sulfate containing 73% iduronic acid. Those molecules found at highest buoyant density appeared to have additional glycosaminoglycan chains that were shorter. Proteoglycans were also extracted from the pressure-bearing distal region of this tendon, where contents of proteoglycan per wet weight of tissue were 3-fold higher and as much as 50% of this was as large as the large proteoglycans from the proximal tissue. Preparations of large proteoglycans from both tendon regions contained molecules capable of interacting with hyaluronic acid.
用4M盐酸胍可从成年牛肌腱近端的纤维部分以较高产率提取蛋白聚糖。它们占该组织干重的比例不到1%。通过氯化铯密度梯度离心、凝胶色谱和离子交换色谱，从其他组织蛋白中分离并纯化出了两类蛋白聚糖。一类是在氯化铯中具有高浮力密度的大型硫酸软骨素蛋白聚糖。通过琼脂糖/聚丙烯酰胺电泳检测，该组分似乎由两到三个亚群组成，尽管无法有效地将它们彼此分离以进行单独表征。总体而言，从Sepharose CL - 2B洗脱的大型蛋白聚糖的洗脱体积分配系数（Kav）为0.1 - 0.5，其核心蛋白的相对分子质量（Mr）大于200,000，谷氨酸、丝氨酸和甘氨酸含量较高。糖胺聚糖链的重均相对分子质量为17,000，且超过98%的糖醛酸为葡萄糖醛酸。该类仅占组织总蛋白聚糖的12%。其余88%的蛋白聚糖似乎代表一类小分子，它们在洗脱体积分配系数（Kav）= 0.70时从Sepharose CL - 2B洗脱。它们在氯化铯梯度中的浮力密度范围为大于或等于1.51至1.30 g/ml。在用软骨素酶ABC消化去除糖胺聚糖链后，其核心蛋白的表观相对分子质量为48,000。该核心蛋白的天冬氨酸/天冬酰胺和亮氨酸含量特别高。糖胺聚糖链的重均相对分子质量为37,000，为含73%艾杜糖醛酸的硫酸皮肤素。在最高浮力密度下发现的那些分子似乎还有额外的较短糖胺聚糖链。也从该肌腱承受压力的远端区域提取了蛋白聚糖，该区域每湿重组织中的蛋白聚糖含量高3倍，其中高达50%与近端组织中的大型蛋白聚糖一样大。来自两个肌腱区域的大型蛋白聚糖制剂都含有能够与透明质酸相互作用的分子。
