Kielty C M, Woolley D E, Whittaker S P, Shuttleworth C A
School of Biological Sciences, University of Manchester, UK.
FEBS Lett. 1994 Aug 29;351(1):85-9. doi: 10.1016/0014-5793(94)00818-3.
We present ultrastructural and biochemical evidence for the turnover of intact fibrillin microfibrils by the serine proteinases, neutrophil elastase, chymotrypsin and trypsin. Rotary shadowing electron microscopy revealed that serine proteinase treatment of intact microfibrils isolated from foetal bovine skin resulted in extensive degradation. Microfibrils were destroyed by neutrophil elastase and effectively disrupted by chymotrypsin and trypsin, with no morphologically identifiable arrays remaining. Evidence of defined fibrillin degradation products was obtained by Western blotting of these enzyme-treated fibrillin assemblies. Fibrillin immunoprecipitated from dermal fibroblast culture medium was also comprehensively degraded by these enzymes. These observations demonstrate that serine proteinases are potent effectors for the physiological and pathological catabolism of microfibrils, and suggest a key role in elastic fibre degradation.
我们提供了超微结构和生化证据，证明丝氨酸蛋白酶（中性粒细胞弹性蛋白酶、胰凝乳蛋白酶和胰蛋白酶）可使完整的原纤维微原纤维发生周转。旋转阴影电子显微镜显示，用丝氨酸蛋白酶处理从胎牛皮肤分离出的完整微原纤维会导致广泛降解。微原纤维被中性粒细胞弹性蛋白酶破坏，并被胰凝乳蛋白酶和胰蛋白酶有效破坏，没有留下形态上可识别的阵列。通过对这些酶处理的原纤维组件进行蛋白质印迹分析，获得了明确的原纤维降解产物的证据。从真皮成纤维细胞培养基中免疫沉淀的原纤维也被这些酶全面降解。这些观察结果表明，丝氨酸蛋白酶是微原纤维生理和病理分解代谢的有效效应物，并提示其在弹性纤维降解中起关键作用。
