N-Terminal Fragment of Vimentin Is Responsible for Binding of Mitochondria In Vitro

IF 1.1 Q4 CELL BIOLOGY

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Pub Date : 2022-06-16 DOI:10.1134/S1990747822030059

A. A. Dayal, N. V. Medvedeva, A. A. Minin

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Abstract—

The role of intermediate filaments in the regulation of mitochondrial functions has become evident from recent studies. For example, vimentin has been shown to affect mitochondrial motility and the level of their membrane potential. However, the mechanism of their interaction is still largely unexplored. In particular, it is unknown whether vimentin can bind directly to mitochondria or whether any intermediate proteins are needed. In this study, using bioinformatics tools, we show that the vimentin sequence has a region in the N-terminal domain, which can play the role of a mitochondrial targeting peptide that probably directs vimentin to mitochondria and causes its binding with these organelles. In order to test this possibility, the binding of mitochondria isolated from rat liver with protofilaments formed by human recombinant vimentin was investigated using centrifugation through sucrose “cushion”. We demonstrate that vimentin can bind to mitochondria in vitro. We also show that the action of a mitochondrial protease leads to the loss of the N-terminal part of the vimentin molecule and its interaction with mitochondria is disrupted. Inhibitory analysis revealed that the atypical calpain, a cysteine Ca²⁺-dependent protease that is insensitive to the inhibitor calpastatin, is responsible for its degradation.

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Vimentin n端片段在体外与线粒体结合

最近的研究表明，中间丝在线粒体功能调控中的作用已经很明显。例如，vimentin已被证明影响线粒体运动和膜电位水平。然而，它们相互作用的机制在很大程度上仍未被探索。特别是，尚不清楚vimentin是否可以直接与线粒体结合，或者是否需要任何中间蛋白。在这项研究中，我们使用生物信息学工具，发现vimentin序列在n端区域有一个区域，该区域可以发挥线粒体靶向肽的作用，可能将vimentin引导到线粒体并导致其与这些细胞器结合。为了验证这种可能性，我们用蔗糖“缓冲垫”离心研究了从大鼠肝脏分离的线粒体与人重组vimentin形成的原丝的结合。我们在体外证明了波形蛋白可以与线粒体结合。我们还表明，线粒体蛋白酶的作用导致静脉蛋白分子n端部分的丢失，其与线粒体的相互作用被破坏。抑制分析显示，非典型钙蛋白酶，半胱氨酸Ca2+依赖性蛋白酶，是不敏感的抑制剂钙pastatin，负责其降解。

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来源期刊

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology CELL BIOLOGY-

CiteScore

1.40

自引率

0.00%

发文量

期刊介绍： Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology is an international peer reviewed journal that publishes original articles on physical, chemical, and molecular mechanisms that underlie basic properties of biological membranes and mediate membrane-related cellular functions. The primary topics of the journal are membrane structure, mechanisms of membrane transport, bioenergetics and photobiology, intracellular signaling as well as membrane aspects of cell biology, immunology, and medicine. The journal is multidisciplinary and gives preference to those articles that employ a variety of experimental approaches, basically in biophysics but also in biochemistry, cytology, and molecular biology. The journal publishes articles that strive for unveiling membrane and cellular functions through innovative theoretical models and computer simulations.