Bullimore D W, Phelps C F, Jayson M I
Biochem J. 1981 Nov 1;199(2):433-9. doi: 10.1042/bj1990433.
Proteoglycans were prepared from human femoral-head articular cartilage by using either guanidinium hydrochloride or MgCl2 as extractant, followed by density-gradient centrifugation. The proteoglycan subunit had a particle weight of 2.6 x 10(6), with a radius of gyration, RG, of 68.5 nm in 150 mM-NaCl/20 mM-sodium phosphate buffer, pH 7.0. The proteoglycan aggregate had a particle weight of 3.7 x 10(6) (RG 84 nm) for guanidinium hydrochloride extracts and 8.7 x 10(6) (RG 118 nm) for MgCl2 extracts in the same buffer. The addition of excess of high-molecular-weight hyaluronate did not significantly alter the particle size of the aggregate. The small increase in size probably reflects a rapid equilibrium between hyaluronate and proteoglycan monomers, and is not due to proteolytic cleavage producing non-aggregating units. Experiments that support the rapid-interaction hypothesis include analytical ultracentrifugation and column chromatography. This interaction does not appear to be pressure-sensitive at 20 degrees C, but is sensitive to temperature variation near the physiological range.
采用盐酸胍或氯化镁作为提取剂,从人股骨头关节软骨中制备蛋白聚糖,随后进行密度梯度离心。在150 mM氯化钠/20 mM磷酸钠缓冲液(pH 7.0)中,蛋白聚糖亚基的分子量为2.6×10⁶,旋转半径RG为68.5 nm。在相同缓冲液中,盐酸胍提取物的蛋白聚糖聚集体分子量为3.7×10⁶(RG 84 nm),氯化镁提取物的为8.7×10⁶(RG 118 nm)。添加过量的高分子量透明质酸不会显著改变聚集体的粒径。粒径的小幅增加可能反映了透明质酸与蛋白聚糖单体之间的快速平衡,而非蛋白水解产生非聚集单元所致。支持快速相互作用假说的实验包括分析超速离心和柱色谱法。在20℃时,这种相互作用似乎对压力不敏感,但对生理范围附近的温度变化敏感。