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Translocation of Paratropomyolsin from the A-I Junctiion Region to Contribute the Resolution of Rigor-Mortis of Postmortem Skeletal Muscles

副肌醇蛋白从 A-I 交界区的易位有助于解决死后骨骼肌的尸僵

基本信息

批准号：
12660244
负责人：
YAMANOUE Minoru
金额：
$ 2.3万
依托单位：
KOBE UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660244/
关键词：
Meat Tenderizatiion Ageing Paratropomyosin Connectin Saromere A-I junction 2D-PAGE A-I接合部

项目摘要

We have suggested that myofibrillar protein, paratropomyosin, is bound to connectin in the A-I junction region of a saroomere to contribute to meat tenderization in postmortem ageing. This study was conducted to determine the paratropomyosin binding site on connectin in living and immediately postmortem chicken muscles in order to confirm the localization of paratropomyosin on connectin filaments, and to clarify the mechanism for release ofparatropomyosin from the site by the high Ca^<2+> concentration.β-Connectin, which was a native form of connectin, was purified from chicken breast muscle and digested by protease under various conditions. The digested β-connectin fragments were separated by two-dimensional polyacrylamide gel electrophoresis. The 43-kDa fragment, to which paratropomyosin bound at the A-I junction region, was detected by biotinylated paratropomyosiii and anti-connectin monoclonal antibody The N-terminal sequence of the 43-kDa fragment was found to be YQFRVYAVNK, similar to the sequence of 7556-7565 amino acids in the 151 fibronectin type 3 domain that was located at the A-I junction region of human cardiac titin/connectin. Therefore, we propose that paratropomyosin binds to the 43-kDa fragment from j3-connectin at the A-I junction region in both living muscle and in muscle immediately postmortem, and the N-terminus of the 43-kDa fragment is localized in the 151 domain. Moreover, we have purified the 400-kDa fragment from chicken myofibrils, which is a part of N-terminal side of p-connectin and contains the 43-kDa fragment in its structure. The analysis of the primary structure of the 400-kDa fragment is in progress to specify the paratropomyosin binding site on connectin.

我们已经提出，肌原纤维蛋白（paratropomyosin）与肉粒a - i连接区域的连接蛋白结合，在死后老化过程中促进肉的嫩化。本研究通过测定活鸡和死后鸡肌肉中连接蛋白上的副atropomyosin结合位点，以确定其在连接蛋白细丝上的定位，并阐明高Ca^<2+>浓度下副atropomyosin从该位点释放的机制。β-连接蛋白是一种天然形式的连接蛋白，从鸡胸肌中分离得到，并在不同条件下用蛋白酶消化。通过双向聚丙烯酰胺凝胶电泳分离酶切后的β-连接蛋白片段。用生物素化的paratropomyosiii和抗连接蛋白单克隆抗体检测了A-I连接区paratropomyosin结合的43-kDa片段，发现43-kDa片段的n端序列为YQFRVYAVNK，与位于人心脏titin/connectin A-I连接区的151型纤维连接蛋白3型结构域的7556-7565个氨基酸序列相似。因此，我们认为paratropomyosin在活肌和死后肌肉的A-I连接区与j3-connectin的43-kDa片段结合，并且43-kDa片段的n端位于151结构域。此外，我们从鸡肌原纤维中纯化了400 kda片段，该片段是p-连接蛋白n端的一部分，其结构中含有43 kda片段。目前正在对400-kDa片段的初级结构进行分析，以确定连接蛋白上的paratropomyosin结合位点。