Dawson P J, Hulme J S, Lloyd C W
J Cell Biol. 1985 May;100(5):1793-8. doi: 10.1083/jcb.100.5.1793.
The monoclonal antibody (anti-IFA) raised (Pruss et al., 1981, Cell 27:419-428) against an intermediate filament antigen, which is widespread throughout phylogeny, has been shown here to cross-react with higher plants. On immunoblotting, anti-IFA cross-reacted with proteins in homogenates of carrot suspension cells and of meristematic cells from onion root tips. A 50-kD cross-reactive protein was enriched in a fraction that consisted of detergent-insoluble bundles of 7-nm fibrils from carrot protoplasts (Powell et al., 1982, J. Cell Sci. 56:319-335). By use of indirect immunofluorescence, anti-IFA stained formaldehyde-fixed onion meristematic cells and carrot protoplasts in patterns approximating those obtained with monoclonal anti-tubulins. That anti-IFA was not recognizing plant tubulins was established by use of immunoblots of two-dimensional gels on which the proteins that comprised isolated fibrillar bundles and taxol-purified carrot tubulins had been separated. The two groups of proteins had different positional coordinates: anti-IFA recognized the fibrillar bundle proteins, and anti-tubulins recognized plant microtubule proteins with no cross-reaction to the heterologous proteins. Likewise, formaldehyde-fixed taxol microtubules from carrot cells could be stained with anti-tubulin but not with anti-IFA. It is concluded that an epitope common to intermediate filaments from animals co-distributes with microtubules in higher plant cells.
针对一种在整个系统发育过程中广泛存在的中间丝抗原产生的单克隆抗体(抗IFA)(Pruss等人,1981年,《细胞》27:419 - 428),在此已显示与高等植物发生交叉反应。在免疫印迹中,抗IFA与胡萝卜悬浮细胞和洋葱根尖分生细胞匀浆中的蛋白质发生交叉反应。一种50-kD的交叉反应蛋白在一个组分中富集,该组分由来自胡萝卜原生质体的7纳米纤维状去污剂不溶性束组成(Powell等人,1982年,《细胞科学杂志》56:319 - 335)。通过间接免疫荧光法,抗IFA以与单克隆抗微管蛋白所获得的模式近似的方式对甲醛固定的洋葱分生细胞和胡萝卜原生质体进行染色。通过对二维凝胶的免疫印迹确定抗IFA不识别植物微管蛋白,在二维凝胶上已分离出构成分离的纤维状束的蛋白质和紫杉醇纯化的胡萝卜微管蛋白。这两组蛋白质具有不同的位置坐标:抗IFA识别纤维状束蛋白,抗微管蛋白识别植物微管蛋白且与异源蛋白无交叉反应。同样,来自胡萝卜细胞的甲醛固定的紫杉醇微管可用抗微管蛋白染色,但不能用抗IFA染色。得出的结论是,动物中间丝共有的一个表位与高等植物细胞中的微管共同分布。