Cross-reactivity of antibodies specific for flagellar tektin and intermediate filament subunits.
Cross-reactivity of antibodies specific for flagellar tektin and intermediate filament subunits.
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DOI：
10.1083/jcb.104.6.1563
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发表时间：
1987-06
期刊：
影响因子：
--
通讯作者：
Piperno G
中科院分区：
文献类型：
--
作者：
Chang XJ;Piperno G
制备了对每种鞭毛tektins具有特异性的单克隆抗体，并用于确定缺乏纤毛或鞭毛的细胞中是否存在类似于tektin细丝的结构。该分析通过几种细胞系的双标记免疫荧光显微镜和蛋白质组分的免疫印迹进行。四种抗-tektin抗体中的两种，抗体3-7-1和3-10-1，其结合C-tektin的不同表位，标记3 T3、HeLa、PtK 2和BHK-21细胞以及肌管。抗体3-7-1染色细胞中的中间丝结构并结合细胞骨架蛋白制剂中的波形蛋白或结蛋白;而抗体3-10-1染色细胞中的核膜并结合细胞骨架蛋白或核纤层制剂中的核纤层蛋白A和C。C-tektin和中间丝蛋白之间的结构相似性可能扩展到两个以上的表位，因为多克隆抗体抗波形蛋白和抗结蛋白结合C-tektin。这些多克隆抗体也与A-tektin结合。单克隆和多克隆抗体与tektin和中间丝组分中的表位结合的交叉反应以及tektin亚基中高含量的α-螺旋结构的存在（Linck，R. W.，和G. L. Langevin，1982，J. Cell Sci.，58：1-22）表明tektin和中间丝在其结构的几个部分是同源的。
Monoclonal antibodies specific for each of the flagellar tektins were prepared and used to determine whether structures similar to tektin filaments are present in cells lacking cilia or flagella. This analysis was performed by double-label immunofluorescence microscopy of several cell lines and by immunoblots of protein fractions. Two of the four anti-tektin antibodies, the antibodies 3-7-1 and 3-10-1, which bind different epitopes of the C-tektin, label 3T3, HeLa, PtK2, and BHK-21 cells as well as myotubes. The antibody 3-7-1 stains intermediate filament structures in the cells and binds vimentin or desmin in preparations of cytoskeletal proteins; whereas the antibody 3-10-1 stains nuclear envelopes in the cells and binds lamin A and C in preparations of cytoskeletal proteins or nuclear lamina. Structural similarities between the C-tektin and intermediate filament proteins probably are extended to more than two epitopes because polyclonal antibodies anti-vimentin and anti-desmin bind to C-tektin. These polyclonal antibodies also bind to A-tektin. The cross-reaction of monoclonal and polyclonal antibodies binding to epitopes in tektin and intermediate filament components and the existence of a high content of alpha-helical structure in the tektin subunits (Linck, R. W., and G. L. Langevin, 1982, J. Cell Sci., 58:1-22) indicate that tektin and intermediate filaments are homologous in several parts of their structure.