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针对鞭毛纤丝蛋白和中间丝亚基的特异性抗体的交叉反应性。

Cross-reactivity of antibodies specific for flagellar tektin and intermediate filament subunits.

作者信息

Chang X J, Piperno G

出版信息

J Cell Biol. 1987 Jun;104(6):1563-8. doi: 10.1083/jcb.104.6.1563.

Abstract

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.

摘要

制备了针对每种鞭毛轴纤丝蛋白的单克隆抗体,并用于确定在缺乏纤毛或鞭毛的细胞中是否存在类似于轴纤丝蛋白丝的结构。通过对几种细胞系进行双标免疫荧光显微镜分析以及对蛋白质组分进行免疫印迹来进行此分析。四种抗轴纤丝蛋白抗体中的两种,即结合C轴纤丝蛋白不同表位的抗体3-7-1和3-10-1,可标记3T3、HeLa、PtK2和BHK-21细胞以及肌管。抗体3-7-1可对细胞中的中间丝结构进行染色,并在细胞骨架蛋白制剂中与波形蛋白或结蛋白结合;而抗体3-10-1可对细胞中的核膜进行染色,并在细胞骨架蛋白制剂或核纤层中与核纤层蛋白A和C结合。C轴纤丝蛋白与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,因为抗波形蛋白和抗结蛋白的多克隆抗体可与C轴纤丝蛋白结合。这些多克隆抗体也可与A轴纤丝蛋白结合。单克隆和多克隆抗体与轴纤丝蛋白和中间丝组分中的表位结合的交叉反应以及轴纤丝蛋白亚基中高含量α螺旋结构的存在(林克,R.W.,和G.L.兰热万,1982,《细胞科学杂志》,58:1 - 22)表明轴纤丝蛋白和中间丝在其结构的几个部分是同源的。

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