Silva L C, Borojevic R, Mourão P A
Departamento de Bioquímica, Universidade Federal do Rio de Janeiro, Brazil.
Biochim Biophys Acta. 1992 Jun 9;1139(1-2):96-104. doi: 10.1016/0925-4439(92)90088-5.
Proteoglycans synthesized in vitro by periovular granulomas isolated from livers of schistosome-infected mice were compared with those produced by granuloma-derived cell lines: the primary cell line GR and the permanent cell line GRX. Proteoglycans were metabolically labelled with 35S-sulfate and extracted with 4 M guanidine-HCl containing 2.0% Triton X-100, in the presence of proteinase inhibitors. The radiolabelled proteoglycans were purified and characterized by anion-exchange, gel-filtration and affinity-column chromatography. Heparan sulfate proteoglycans (HS-PGs) and chondroitin sulfate/dermatan sulfate-containing proteoglycans (CS/DS-PGs) were detected in both the culture medium and the cell-associated fractions obtained from GR cells. More than 90% of the cell-associated HS-PG from these cells contained a hydrophobic portion, as evidenced by their ability to bind to octyl-Sepharose. In contrast, among the secreted proteoglycans, it was the CS/DS-PG and not the HS-PG that bound to this resin. The major fractions of cell-associated and secreted proteoglycans from GRX cells were HS-PGs. Similar to HS-PGs from GR cells, 50% of the cell-associated HS-PG bound to octyl-Sepharose, while only 20% of secreted proteoglycans (HS-PGs) bound to this resin. The proteoglycans purified from the whole granuloma were composed mainly of DS-PG, of a size and hydrophobicity similar to the CS/DS-PG from GR cells. Possible correlations among the structure, secretion, distribution and function of proteoglycans in granulomatous reactions are discussed.
将从感染血吸虫小鼠肝脏中分离出的卵泡周围肉芽肿体外合成的蛋白聚糖,与肉芽肿衍生细胞系(原代细胞系GR和永久细胞系GRX)产生的蛋白聚糖进行比较。蛋白聚糖用35S-硫酸盐进行代谢标记,并在蛋白酶抑制剂存在的情况下,用含有2.0% Triton X-100的4M盐酸胍提取。放射性标记的蛋白聚糖通过阴离子交换、凝胶过滤和亲和柱色谱进行纯化和表征。在GR细胞的培养基和细胞相关组分中均检测到硫酸乙酰肝素蛋白聚糖(HS-PGs)和含硫酸软骨素/硫酸皮肤素的蛋白聚糖(CS/DS-PGs)。这些细胞中超过90%的细胞相关HS-PG含有疏水部分,这可通过它们与辛基琼脂糖结合的能力得到证明。相比之下,在分泌的蛋白聚糖中,与该树脂结合的是CS/DS-PG而非HS-PG。GRX细胞的细胞相关和分泌蛋白聚糖的主要部分是HS-PGs。与GR细胞的HS-PGs相似,50%的细胞相关HS-PG与辛基琼脂糖结合,而只有20%的分泌蛋白聚糖(HS-PGs)与该树脂结合。从整个肉芽肿中纯化的蛋白聚糖主要由DS-PG组成,其大小和疏水性与GR细胞的CS/DS-PG相似。讨论了肉芽肿反应中蛋白聚糖的结构、分泌、分布和功能之间可能的相关性。