Savill C M, Ayad S R
Anticancer Res. 1986 Mar-Apr;6(2):321-6.
Fibronectin was partially digested using the lysosomal proteinase cathepsin D and the fragments produced tested for their effect on the rate of DNA synthesis in quiescent Chinese Hamster Fibroblasts (CH23). A mitogenically active digest was produced after digestion of fibronectin with cathepsin D for 2 to 3h at a 1:200 (w/w) ratio of enzyme to substrate at 37 degrees C. Affinity chromatography of the active digest on both gelatin and heparin 4BCl columns demonstrated that the active fragment was gelatin-nonbinding but heparin-binding, and therefore derived from one of the two or three heparin binding domains of intact fibronectin. The active fragment was found to have a Mr35,000 by gel filtration on sephacryl S200. A dose-response curve of the mitogenic fragment purified by gel filtration against DNA synthesis showed it to be maximally active at a concentration of approximately 10 micrograms/ml.
使用溶酶体蛋白酶组织蛋白酶D对纤连蛋白进行部分消化，并测试产生的片段对静止的中国仓鼠成纤维细胞（CH23）中DNA合成速率的影响。在37℃下，以酶与底物1:200（w/w）的比例用组织蛋白酶D消化纤连蛋白2至3小时后，产生了具有促有丝分裂活性的消化产物。在明胶和肝素4BCl柱上对活性消化产物进行亲和层析表明，活性片段不与明胶结合但与肝素结合，因此源自完整纤连蛋白的两个或三个肝素结合结构域之一。通过在Sephacryl S200上进行凝胶过滤发现活性片段的Mr为35,000。通过凝胶过滤纯化的促有丝分裂片段针对DNA合成的剂量反应曲线表明，其在浓度约为10微克/毫升时具有最大活性。
