Stöcker G, Drzeniek Z, Just U, Ostertag W, Siebertz B, Greiling H, Haubeck H D
Institute for Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Aachen, Federal Republic of Germany.
Biochem J. 1996 Jul 1;317 ( Pt 1)(Pt 1):203-12. doi: 10.1042/bj3170203.
Proteoglycans of bone-marrow stromal cells and their extracellular matrix are important components of the microenvironment of haematopoietic tissues. Proteoglycans might also be involved in the interaction of haematopoietic stem and stromal cells. Recently, several studies have been reported on the proteoglycan synthesis of stromal cells, but little is known about the proteoglycan synthesis of haematopoietic stem or progenitor cells. Here we report on the isolation and characterization of proteoglycans from two haematopoietic progenitor cell lines, the murine FDCP-Mix A4 and the human TF-1 cell line. Proteoglycans were isolated from metabolically labelled cells and purified by several chromatographic steps, including anion-exchange and size-exclusion chromatography. Biochemical characterization was performed by electrophoresis or gel-filtration chromatography before and after digestion with glycosaminoglycan-specific enzymes or HNO2 treatment. Whereas FDCP-Mix A4 cells synthesize a homogeneous chondroitin 4-sulphate proteoglycan, isolation and characterization of proteoglycans from the human cell line TF-1 revealed, that TF-1 cells synthesize, in addition to a chondroitin sulphate proteoglycan, a heparan sulphate proteoglycan as major proteoglycan. For this heparan sulphate proteoglycan a core protein size of approx. 59 kDa was determined. Immunochemical analysis of this heparan sulphate proteoglycan revealed that it is not related to the syndecan family nor to glypican.
骨髓基质细胞及其细胞外基质中的蛋白聚糖是造血组织微环境的重要组成部分。蛋白聚糖也可能参与造血干细胞与基质细胞的相互作用。最近，已有多项关于基质细胞蛋白聚糖合成的研究报道，但对于造血干细胞或祖细胞的蛋白聚糖合成了解甚少。在此，我们报告从小鼠FDCP-Mix A4和人TF-1细胞系这两种造血祖细胞系中分离和鉴定蛋白聚糖的情况。从经代谢标记的细胞中分离蛋白聚糖，并通过包括阴离子交换和尺寸排阻色谱在内的多个色谱步骤进行纯化。在用糖胺聚糖特异性酶消化或亚硝酸处理前后，通过电泳或凝胶过滤色谱进行生化表征。FDCP-Mix A4细胞合成一种均一的硫酸软骨素4-硫酸酯蛋白聚糖，而从人TF-1细胞系中分离和鉴定蛋白聚糖发现，TF-1细胞除了合成一种硫酸软骨素蛋白聚糖外，还合成一种硫酸乙酰肝素蛋白聚糖作为主要的蛋白聚糖。对于这种硫酸乙酰肝素蛋白聚糖，确定其核心蛋白大小约为59 kDa。对这种硫酸乙酰肝素蛋白聚糖的免疫化学分析表明，它与多配体蛋白聚糖家族和磷脂酰肌醇蛋白聚糖均无关。
