Campbell M A, Tester A M, Handley C J, Checkley G J, Chow G L, Cant A E, Winter A D, Cain W E
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Arch Biochem Biophys. 1996 May 15;329(2):181-90. doi: 10.1006/abbi.1996.0207.
Bovine collateral ligament synthesized a 35S-labeled large proteoglycan species which eluted with a Kav of approximately 0.27 on Sepharose CL-2B and contained only chondroitin sulfate chains with a molecular mass of approximately 32 kDa. Fluorography of the 35S-labeled core proteins derived from the large ligament proteoglycan revealed a broad range of molecular masses above approximately 200 kDa, which was of comparable size to the four major endogenous core protein bands derived from this proteoglycan detected with 5/6/3-B-3, an antibody directed against terminal unsaturated chondroitin-6-sulfate disaccharides. The core proteins derived from the large ligament proteoglycan exhibited immunoreactivity of 12/21/1-C-6, an antibody specific for a peptide epitope common to both the G1 and G2 domains of aggrecan. Four major core protein bands with molecular masses greater than approximately 200 kDa derived from the large ligament proteoglycan, were detected using the antibodies raised against versican from bovine aorta or human fibroblasts. Compared with aggrecan, the 35S-labeled large ligament proteoglycan was distributed over a broader range of buoyant densities in an associative caesium chloride density gradient. This polydispersity may be indicative of differences in the degree of glycosylation as well as heterogeneity in the size of the large ligament proteoglycan core proteins. The 35S-labeled large ligament proteoglycan also demonstrated the ability to form complexes with an aggrecan aggregate preparation, the majority of which could not be dissociated by the presence of HA10-50. These findings indicate that the large chondrotin sulfate proteoglycan synthesized by bovine collateral ligament may be a versican-like proteoglycan which exhibited the potential to form like protein-stabilized complexes.
牛侧副韧带合成了一种35S标记的大蛋白聚糖，其在琼脂糖CL - 2B上以约0.27的Kav洗脱，并且仅含有分子量约为32 kDa的硫酸软骨素链。对源自大韧带蛋白聚糖的35S标记核心蛋白进行荧光显影，结果显示分子量在约200 kDa以上有一个广泛的范围，其大小与用5/6/3 - B - 3（一种针对末端不饱和硫酸软骨素 - 6 - 硫酸二糖的抗体）检测到的源自该蛋白聚糖的四个主要内源性核心蛋白条带相当。源自大韧带蛋白聚糖的核心蛋白表现出12/21/1 - C - 6的免疫反应性，12/21/1 - C - 6是一种对聚集蛋白聚糖的G1和G2结构域共有的肽表位具有特异性的抗体。使用针对牛主动脉或人成纤维细胞中的多功能蛋白聚糖产生的抗体，检测到源自大韧带蛋白聚糖的四个分子量大于约200 kDa的主要核心蛋白条带。与聚集蛋白聚糖相比，35S标记的大韧带蛋白聚糖在氯化铯关联密度梯度中分布在更宽的浮力密度范围内。这种多分散性可能表明糖基化程度的差异以及大韧带蛋白聚糖核心蛋白大小的异质性。35S标记的大韧带蛋白聚糖还表现出与聚集蛋白聚糖聚集体制剂形成复合物的能力，其中大多数复合物不会因HA10 - 50的存在而解离。这些发现表明，牛侧副韧带合成的大硫酸软骨素蛋白聚糖可能是一种类似多功能蛋白聚糖的蛋白聚糖，其具有形成类似蛋白质稳定复合物的潜力。
