Böttger V, Stasiak P C, Harrison D L, Mellerick D M, Lane E B
Department of Anatomy and Physiology, University of Dundee, Scotland.
Eur J Biochem. 1995 Jul 15;231(2):475-85. doi: 10.1111/j.1432-1033.1995.tb20721.x.
To generate tools for monitoring processing and folding in keratin intermediate filaments, a group of monoclonal antibodies reacting with the intermediate filament protein keratin 19 were studied using different approaches to define the structure and localization of their epitopes. The binding pattern to bacterially expressed human keratin 19 fragments allowed the definition of minimal amino acid sequences required for antibody binding. The screening of overlapping 15-residue peptides confirmed and further specified the epitope locations for a subset of the tested antibodies. In addition, the epitope of an antibody with apparent species-restricted specificity (LE64) was revealed by isolating and characterizing a full-length keratin 19 clone from a PtK2 cDNA library. Taken together with species cross-reactivity of individual antibodies and sequence information obtained by probing a phage display library, specific amino acid residues could be highlighted as likely to be involved in the antibody binding.
为了生成用于监测角蛋白中间丝加工和折叠的工具，研究了一组与中间丝蛋白角蛋白19反应的单克隆抗体，采用不同方法来确定其表位的结构和定位。与细菌表达的人角蛋白19片段的结合模式使得能够确定抗体结合所需的最小氨基酸序列。对重叠的15个残基肽段的筛选证实并进一步明确了一部分测试抗体的表位位置。此外，通过从PtK2 cDNA文库中分离和鉴定全长角蛋白19克隆，揭示了一种具有明显物种限制特异性的抗体（LE64）的表位。结合各抗体的物种交叉反应性以及通过探测噬菌体展示文库获得的序列信息，特定的氨基酸残基可能被突出显示为可能参与抗体结合。
