A kinetic study of salt-induced denaturation of actin during storage at low temperature

低温储存期间盐诱导肌动蛋白变性的动力学研究

基本信息

批准号：
05660307
负责人：
IKEUCHI Yoshihide
金额：
$ 1.34万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660307/
关键词：
Actin Tropomyosin HMM Denaturation Curing DNAase I DNaseI 食肉タンパク質反応速度論

项目摘要

The small amount of F-actomyosin complex acts as a cross-linker with the free myosin molecule on heating, and it is considered to be a prerequisite for actin-induced improvement in the gel formability of myosin. Therefore, the property of heat-induced gel of myosin depends on the stability of actin during treatment with salt. The present work was conducted to elucidate the mechanism of salt-induced denaturation of actin during incubation at a low temperature.1. A kinetic analysis by measuring DNase l inhibition capacity of actin demonstrated that the denaturation of actin obeys the theory of an irreversible continuous reaction (F-ADP-actin ---> G-ADP-actin ---> denatured actin) when actin solution is incubated at a low temperature (0ﾟC).2. The addition of a sufficient amount of ATP to an F-actin solution effectively retards the progress of the denaturation of actin. ATP is thought to stabilize the structure of G-actin depolymerized during treatment with salt. That is, the present of ATP retards the process of G-ADP-actin -->denatured actin.3. It has become apparent that a small amount of HMM accelerates the rate of depolymerization of F-actin during incubation at 0ﾟC.As a result, released G-actin is presumed to enter quickly the denaturation process without ATP.4. When tropomyosin was added to solution containing actin alone or actin-HMM complex, the denaturation of actin was suppressed remarkably between 0.2 M KCI and 0.6 M KCI.From this result, it is clear that tropomyosin stabilizes the actin filament against disassembly at ionic strengths lower than 0.6 M.KCI

少量的F-肌球蛋白复合物充当加热时游离肌球蛋白分子的交联，被认为是肌动蛋白诱导的肌球蛋白凝胶表现可提高的先决条件。因此，热诱导的肌球蛋白凝胶的特性取决于用盐治疗过程中肌动蛋白的稳定性。目前的工作是为了阐明在低温下孵育过程中盐诱导的肌动蛋白变性的机理。1。通过测量肌动蛋白的DNase L抑制能力的动力学分析表明，肌动蛋白的变性服从不可逆的连续反应理论（F-ADP-肌动蛋白----> G-ADP-肌动蛋白 - > G-ADP-肌动蛋白---->变性肌动蛋白）在低温下（0ﾟC）.2 .2。在F-肌动蛋白溶液中添加足够量的ATP有效地阻碍了肌动蛋白变性的进展。人们认为ATP可以稳定用盐处理期间的G-肌动蛋白解聚的结构。也就是说，ATP的当下降低了G-ADP-肌动蛋白的过程 - >变性肌动蛋白3。显然，少量的HMM在0°C孵育过程中加速F-肌动蛋白的解聚速率。结果，释放的G-肌动蛋白会迅速输入不含ATP.4的变性过程。当将真菌素添加到单独包含肌动蛋白或肌动蛋白-HMM复合物的溶液中时，肌动蛋白的变性在0.2 M KCI和0.6 M KCI之间被明显抑制。从该结果中，很明显，Tropomyosin稳定肌动蛋白丝在低于0.6 M.KCCI的脱节强度下稳定肌动蛋白丝。