Tissue-specific interactions of TNI isoforms with other TN subunits and tropomyosins in C-elegans:: The role of the C- and N-terminal extensions
Tissue-specific interactions of TNI isoforms with other TN subunits and tropomyosins in C-elegans:: The role of the C- and N-terminal extensions
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DOI：
10.1016/j.bbapap.2007.01.003
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发表时间：
2007-04-01
影响因子：
3.2
通讯作者：
Kagawa, Hiroaki
中科院分区：
文献类型：
--
作者：
Amin, Md. Ziaul;Bando, Tetsuya;Kagawa, Hiroaki
本研究的目的是研究秀丽隐杆线虫体壁肌肉特有的三种肌钙蛋白I亚型的C端延伸的功能，并了解肌钙蛋白I与肌钙蛋白C/T和原肌球蛋白在TN复合体内的分子相互作用。我们构建了多个表达载体，在大肠杆菌中生成了三个体壁和一个咽肌钙蛋白I亚型的重组蛋白。使用抗体进行蛋白覆盖试验和Western blot分析。我们证明咽部的TNI-4仅与肌钙蛋白C/T和原肌球蛋白的咽部亚型相互作用。相反，体壁的TNI-2结合了这些成分的体壁和咽异构体。与其他无脊椎动物类似，肌钙蛋白I的n端参与与肌钙蛋白c的相互作用。全长肌钙蛋白I是与原肌球蛋白同型体相互作用所必需的。C端延伸的缺失对体壁肌钙蛋白I与其他肌细丝肌钙蛋白C/T和原肌球蛋白同型体的结合没有直接影响。(C) 2007 Elsevier B.V.版权所有
The aim of this study is to investigate the function of the C-terminal extension of three troponin I isoforms, that are unique to the body wall muscles of Caenorhabditis elegans and to understand the molecular interactions within the TN complex between troponin I with troponin C/T, and tropomyosin. We constructed several expression vectors to generate recombinant proteins of three body wall and one pharyngeal troponin I isoforms in Escherichia coli. Protein overlay assays and Western blot analyses were pet-formed using antibodies. We demonstrated that pharyngeal TNI-4 interacted with only the pharyngeal isoforms of troponin C/T and tropomyosin. In contrast, the body wall TNI-2 bound both the body wall and pharyngeal isoforms of these components. Similar to other invertebrates, the N-terminus of troponin I contributes to interactions with troponin C. Full-length troponin I was essential for interactions with tropomyosin isoforms. Deletion of the C-terminal extension had no direct effect on the binding of the body wall troponin I to other muscle thin filament troponin C/T and tropomyosin isoforms. (C) 2007 Elsevier B.V. All rights reserved.