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首页> 外文期刊>Cell Division >Epitope of titin A-band-specific monoclonal antibody Tit1 5 H1.1 is highly conserved in several Fn3 domains of the titin molecule. Centriole staining in human, mouse and zebrafish cells
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Epitope of titin A-band-specific monoclonal antibody Tit1 5 H1.1 is highly conserved in several Fn3 domains of the titin molecule. Centriole staining in human, mouse and zebrafish cells

机译：titin A波段特异性单克隆抗体Tit1 5 H1.1的表位在titin分子的几个Fn3结构域中高度保守。人类，小鼠和斑马鱼细胞中的离心染色

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Background Previously we have reported on the development of a new mouse anti-titin monoclonal antibody, named MAb Titl 5 H1.1, using the synthetic peptide N-AVNKYGIGEPLESDSVVAK-C which corresponds to an amino acid sequence in the A-region of the titin molecule as immunogen. In the human skeletal muscles, MAb Titl 5 H1.1 reacts specifically with titin in the A-band of the sarcomere and in different non-muscle cell types with nucleus and cytoplasm, including centrioles. In this report we have studied the evolutionary aspects of the binding of MAb Tit1 5 H1.1 with its target antigen (titin). Results We have specified the epitope area of MAb Tit1 5 H1.1 by subpeptide mapping to the hexapeptide N-AVNKYG-C. According to protein databases this amino acid sequence is located in the COOH-terminus of several different Fn3 domains of the A-region of titin molecule in many organisms, such as human being, mouse, rabbit, zebrafish (Danio rerio), and even in sea squirt (Ciona intestinalis). Our immunohisto- and cytochemical studies with MAb Tit1 5 H1.1 in human, mouse and zebrafish tissues and cell cultures showed a striated staining pattern in muscle cells and also staining of centrioles, cytoplasm and nuclei in non-muscle cells. Conclusions The data confirm that titin can play, in addition to the known roles in striated muscle cells also an important role in non-muscle cells as a centriole associated protein. This phenomenon is highly conserved in the evolution and is related to Fn3 domains of the titin molecule. Using titin A-band-specific monoclonal antibody MAb Tit1 5 H1.1 it was possible to locate titin in the sarcomeres of skeletal muscle cells and in the centrioles, cytoplasm and nuclei of non-muscle cells in phylogenetically so distant organisms as Homo sapiens, Mus musculus and zebrafish (Danio rerio).
机译：背景技术先前我们曾报道过使用合成肽N-AVNKYGIGEPLESDSVVAK-C开发名为Mab Titl 5 H1.1的小鼠抗titin单克隆抗体的新方法，该肽对应于titin A区的氨基酸序列。分子作为免疫原。在人的骨骼肌中，MAb Titl 5 H1.1与肌节A带中的肌动蛋白发生特异性反应，并在具有核和细胞质（包括中心粒）的不同非肌肉细胞类型中发生特异性反应。在本报告中，我们研究了MAb Tit1 5 H1.1与其靶抗原（titin）结合的进化方面。结果我们通过亚肽定位到六肽N-AVNKYG-C来确定MAb Tit1 5 H1.1的表位区域。根据蛋白质数据库，该氨基酸序列位于许多生物体（如人，小鼠，兔，斑马鱼（斑马鱼），甚至在人类中）中titin分子A区几个不同Fn3结构域的COOH末端。海鞘（Ciona intestinalis）。我们在人，小鼠和斑马鱼组织和细胞培养物中对MAb Tit1 5 H1.1进行的免疫组织化学和细胞化学研究显示，肌肉细胞中有横纹状染色，非肌肉细胞中也有中心粒，细胞质和细胞核染色。结论数据证实，除了在纹状肌细胞中的已知作用外，titin还能发挥作用，作为中心蛋白相关蛋白在非肌肉细胞中也起重要作用。这种现象在进化过程中高度保守，与titin分子的Fn3结构域有关。使用titin A波段特异性单克隆抗体MAb Tit1 5 H1.1，可以将titin定位于骨骼肌细胞的肉瘤中以及系统发育如此遥远的有机体（如智人）的中心，非肌细胞的细胞质和细胞核中，小家鼠和斑马鱼（Danio rerio）。

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