Sheehan J K, Nieduszynski I A, Phelps C F
Biochem J. 1978 Apr 1;171(1):109-14. doi: 10.1042/bj1710109.
Proteoglycans from pig laryngeal cartilage prepared by dissociative extraction in guanidine hydrochloride were studied in dilute solution by light-scattering and ultracentrifugation. In buffered 150mM-NaCl, pH7.4, the proteoglycan particle weights were about 5x10(6) daltons, but at 100mM-, 200mM- and 300mM-NaCl particle weights of 2.5x10(6)--3.0x10(6) daltons were observed. These results, together with corroborative evidence from sedimentation-velocity experiments, were interpreted in terms of proteoglycans self-associating at physiological ionic strength. The data were examined by using a proteoglycan monomer-dimer model. Proteoglycan preparations that had thiol groups partially carboxymethylated gave particle weights of 3.2x10(6)--3.5x10(6) daltons in 150mM-NaCl, which suggested that carboxymethylation inhibited multimerization and hence that the protein core is implicated in the binding site. Further studies showed that the multimers were stable to 60 degrees C, unlike the hyaluronate-proteoglycan complex.
对通过在盐酸胍中进行解离提取制备的猪喉软骨蛋白聚糖,在稀溶液中用光散射和超速离心法进行了研究。在pH7.4的150mM - NaCl缓冲液中,蛋白聚糖的颗粒重量约为5×10⁶道尔顿,但在100mM、200mM和300mM - NaCl时,观察到颗粒重量为2.5×10⁶ - 3.0×10⁶道尔顿。这些结果,连同沉降速度实验的佐证证据,被解释为蛋白聚糖在生理离子强度下会自我缔合。使用蛋白聚糖单体 - 二聚体模型对数据进行了检验。硫醇基团部分羧甲基化的蛋白聚糖制剂在150mM - NaCl中颗粒重量为3.2×10⁶ - 3.5×10⁶道尔顿,这表明羧甲基化抑制了多聚化,因此蛋白质核心与结合位点有关。进一步研究表明,与透明质酸 - 蛋白聚糖复合物不同,多聚体在60℃下是稳定的。