Stöcker G, Fischer D C
Department of Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Aachen, F.R.G.
J Chromatogr. 1990 Nov 23;521(2):311-24. doi: 10.1016/0021-9673(90)85055-z.
Two chromatographic procedures for the isolation and purification of proteoglycans (PG) and their related glycosaminoglycan (GAG) peptides are described. PG from human aorta were isolated from tissue extract by sequential ion-exchange, size-exclusion and hydroxyapatite chromatography. Final purification of samples was achieved by chromatography on Mono Q. Homogeneity of samples was demonstrated by Western blot analysis of biotin-labelled compounds prior to and after enzymatic digestion and dual-wavelength detection in size-exclusion chromatography. The purity of samples obtained by the procedure described was sufficient for protein sequence analysis. GAG preparations of bovine trachea cartilage were purified by the sequential use of strong anion-exchange supports. Molecular weight distribution and sensitivity to treatment with glycan-specific enzymes was shown by size-exclusion chromatography.
本文描述了两种用于分离和纯化蛋白聚糖（PG）及其相关糖胺聚糖（GAG）肽的色谱方法。人主动脉中的PG通过离子交换、尺寸排阻和羟基磷灰石色谱法从组织提取物中分离出来。样品的最终纯化通过Mono Q色谱法实现。在酶消化前后，通过对生物素标记化合物进行蛋白质印迹分析以及尺寸排阻色谱法中的双波长检测，证明了样品的均一性。通过所述方法获得的样品纯度足以进行蛋白质序列分析。牛气管软骨的GAG制剂通过依次使用强阴离子交换支持物进行纯化。通过尺寸排阻色谱法显示了分子量分布和对聚糖特异性酶处理的敏感性。
