Kielty C M, Whittaker S P, Shuttleworth C A
Department of Medicine, University of Manchester, UK.
FEBS Lett. 1996 May 20;386(2-3):169-73. doi: 10.1016/0014-5793(96)00423-1.
We have investigated the potential association of proteoglycans with intact fibrillin-containing microfibrils from foetal bovine elastic tissues and with newly synthesised fibrillin in human and bovine cell cultures. Microfibril integrity was disrupted by chondroitinase ABC lyase and chondroitinase AC lyase, but not by keratanase or hyaluronidase. Following chondroitinase treatment, beads were disrupted but the underlying fibrillar scaffold appeared intact. Cuprolinic blue was prominently associated with beaded domains at a critical electrolyte concentration. Electron-dense rods were often associated with cuprolinic blue-treated microfibrils isolated from fixed tissues. Positive staining revealed charged foci at the beads. Newly synthesised fibrillin could be labelled with 35S TransLabel, [3H]glucosamine or 35SO4 but its electrophoretic mobility was not influenced by treatment with chondroitinase ABC or AC lyase. A diffuse 35SO4-labelled chondroitinase-sensitive component with a resistant band (Mr 35000) co-immunoprecipitated with fibrillin. These experiments indicate that chondroitin sulphate proteoglycans associate with fibrillin and contribute to microfibril assembly. This association has major implications for microfibril function in health and disease.
我们研究了蛋白聚糖与来自胎牛弹性组织的完整含原纤蛋白微原纤维以及人与牛细胞培养物中新合成的原纤蛋白之间的潜在关联。微原纤维的完整性被软骨素酶ABC裂解酶和软骨素酶AC裂解酶破坏，但未被角蛋白酶或透明质酸酶破坏。软骨素酶处理后，珠子被破坏，但下面的纤维状支架看起来完好无损。在临界电解质浓度下，铜试剂蓝与珠状结构域显著相关。电子致密棒通常与从固定组织中分离出的经铜试剂蓝处理的微原纤维相关。阳性染色显示珠子处有带电焦点。新合成的原纤蛋白可用35S TransLabel、[3H]葡萄糖胺或35SO4标记，但其电泳迁移率不受软骨素酶ABC或AC裂解酶处理的影响。一种具有抗性条带（Mr 35000）的弥散性35SO4标记的软骨素酶敏感成分与原纤蛋白共免疫沉淀。这些实验表明硫酸软骨素蛋白聚糖与原纤蛋白相关联，并有助于微原纤维的组装。这种关联对健康和疾病中的微原纤维功能具有重要意义。
