Membrane-associated chondroitin sulfate proteoglycans of human lung fibroblasts.
Membrane-associated chondroitin sulfate proteoglycans of human lung fibroblasts.
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DOI：
10.1083/jcb.108.3.1165
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发表时间：
1989-03
期刊：
影响因子：
--
通讯作者：
Van den Berghe H
中科院分区：
文献类型：
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作者：
David G;Lories V;Heremans A;Van der Schueren B;Cassiman JJ;Van den Berghe H
培养的人胎儿肺成纤维细胞产生一些硫酸软骨素蛋白多糖，这些蛋白多糖在含有4 M氯化胍的朝向缓冲液中作为聚集体提取。当Triton X-100加入缓冲液时，聚集的蛋白多糖从Sepharose CL4B和2B中被排除，但以0.53的Kav值从Sepharose CL4B中洗脱出来。相反，一些洗涤剂可提取的硫酸软骨素蛋白聚糖可以并入脂质体，这表明存在疏水膜插层硫酸软骨素蛋白聚糖部分。疏水硫酸软骨素蛋白聚糖的纯化制剂含有90和52 kD的两种主要核心蛋白形式。用疏水蛋白聚糖免疫的BALB/c小鼠的骨髓瘤细胞与脾脏细胞融合获得的单克隆抗体（F58-7D8）可识别90- kd核心蛋白，但不能识别52-kD核心蛋白。抗体识别的表位暴露在培养的人肺成纤维细胞的表面和体内几种基质细胞的表面，也暴露在库普弗细胞和表皮细胞的表面。这些小的膜相关硫酸软骨素蛋白聚糖的核心蛋白可能与先前通过生化、免疫学和分子生物学方法在人成纤维细胞中发现的核心蛋白不同。
Cultured human fetal lung fibroblasts produce some chondroitin sulfate proteoglycans that are extracted as an aggregate in chaotropic buffers containing 4 M guanidinium chloride. The aggregated proteoglycans are excluded from Sepharose CL4B and 2B, but become included, eluting with a Kav value of 0.53 from Sepharose CL4B, when Triton X-100 is included in the buffer. Conversely, some of the detergent-extractable chondroitin sulfate proteoglycans can be incorporated into liposomes, suggesting the existence of a hydrophobic membrane-intercalated chondroitin sulfate proteoglycan fraction. Purified preparations of hydrophobic chondroitin sulfate proteoglycans contain two major core protein forms of 90 and 52 kD. A monoclonal antibody (F58-7D8) obtained from the fusion of myeloma cells with spleen cells of BALB/c mice that were immunized with hydrophobic proteoglycans recognized the 90- but not the 52-kD core protein. The epitope that is recognized by the antibody is exposed at the surface of cultured human lung fibroblasts and at the surface of several stromal cells in vivo, but also at the surface of Kupffer cells and of epidermal cells. The core proteins of these small membrane-associated chondroitin sulfate proteoglycans are probably distinct from those previously identified in human fibroblasts by biochemical, immunological, and molecular biological approaches.