Keski-Oja J, Sen A, Todaro G J
J Cell Biol. 1980 Jun;85(3):527-33. doi: 10.1083/jcb.85.3.527.
Affinity chromatography with actin-Sepharose conjugates of purified human fibronectin, normal human plasma, or serum-free culture fluid from human fibroblasts showed that fibronectin molecules can directly bind to actin. A quantitative recovery of soluble human fibronectin was accomplished by chromatography on actin immobilized on Sepharose beads. Human fibronectin molecules bound to actin-Sepharose were eluted with 0.25--0.35 M potassium bromide, and these molecules competed in a species-specific radioimmunoassay for human fibronectin. The subunits of fibronectin isolated by actin-Sepharose chromatography comigrated in SDS polyacrylamide gel electrophoresis with those of electrophoretically homogeneous fibronectin purified by conventional procedures. The efficient direct binding of fibronectin to actin suggests that interactions between these proteins might also take place in vivo but further studies are needed to elucidate the biological significance of this affinity.
用纯化的人纤连蛋白、正常人血浆或人成纤维细胞的无血清培养液与肌动蛋白-琼脂糖缀合物进行亲和层析表明,纤连蛋白分子可直接与肌动蛋白结合。通过在固定于琼脂糖珠上的肌动蛋白上进行层析,可定量回收可溶性人纤连蛋白。结合到肌动蛋白-琼脂糖上的人纤连蛋白分子用0.25 - 0.35M溴化钾洗脱,并且这些分子在人纤连蛋白的种属特异性放射免疫测定中具有竞争性。通过肌动蛋白-琼脂糖层析分离的纤连蛋白亚基在SDS聚丙烯酰胺凝胶电泳中与通过传统方法纯化的电泳纯纤连蛋白的亚基一起迁移。纤连蛋白与肌动蛋白的有效直接结合表明这些蛋白质之间的相互作用也可能在体内发生,但需要进一步研究来阐明这种亲和力的生物学意义。