Impact of actin glutathionylation on the actomyosin-S1 ATPase.

Impact of actin glutathionylation on the actomyosin-S1 ATPase.
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肌动蛋白谷胱甘肽化对肌动球蛋白-S1 ATP 酶的影响。

DOI:
10.1021/bi900669m
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发表时间:
2009
期刊:
影响因子:
2.9
通讯作者:
Ogut,Ozgur
Ogut,Ozgur
中科院分区:
生物学3区
文献类型:
--
作者:
Pizarro,GresinO;Ogut,Ozgur

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细胞内蛋白质的谷胱甘肽化是蛋白质功能的既定生理调节剂。在多种模型中,包括心脏缺血再灌注,氧化应激增加导致肌节肌动蛋白谷胱甘肽化。我们假设肌动蛋白谷胱甘肽化可能在这种病理生理状态期间观察到的心肌收缩力的多因素变化中发挥作用。因此,研究了谷胱甘肽化肌动蛋白对与肌球蛋白-S1 相互作用的功能影响。用谷胱甘肽化的 F-肌动蛋白替代未修饰的 F-肌动蛋白降低了最大肌动球蛋白-S1 ATP酶,并且伴随着稳态 ATP酶的活化能的增加。稳态结合的测量并未表明肌动蛋白谷胱甘肽化对肌球蛋白-S1 的结合有很大影响。然而,通过停流方法测定的瞬时结合和解离动力学表明,尽管肌动蛋白谷胱甘肽化没有显着改变肌球蛋白-S1结合的速率常数,但在ADP存在的情况下,ATP诱导的肌球蛋白-S1脱离速率显着降低。这些结果表明,肌动蛋白谷胱甘肽化可能在心肌氧化应激后收缩力的改变中发挥有限但明确的作用,特别是通过肌动球蛋白 ATP 酶活性的降低。
Glutathionylation of intracellular proteins is an established physiological regulator of protein function. In multiple models, including ischemia-reperfusion of the heart, increased oxidative stress results in the glutathionylation of sarcomeric actin. We hypothesized that actin glutathionylation may play a role in the multifactorial change in cardiac muscle contractility observed during this pathophysiological state. Therefore, the functional impact of glutathionylated actin on the interaction with myosin-S1 was examined. Substituting glutathionylated F-actin for unmodified F-actin reduced the maximum actomyosin-S1 ATPase, and this was accompanied by an increase in the activation energy of the steady state ATPase. Measurement of steady state binding did not suggest a large impact of actin glutathionylation on the binding to myosin-S1. However, transient binding and dissociation kinetics determined by stopped-flow methods demonstrated that although actin glutathionylation did not significantly alter the rate constant of myosin-S1 binding, there was a significant decrease in the rate of ATP-induced myosin-S1 detachment in the presence of ADP. These results suggest that actin glutathionylation may play a limited but defined role in the alteration of contractility following oxidative stress to the myocardium, particularly through a decrease in the actomyosin ATPase activity.
DOI: 10.1021/bi972851y
发表时间: 1998-05-05
期刊: BIOCHEMISTRY
影响因子: 2.9
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