A Bioresistant Nitroxide Spin Label for In‐Cell EPR Spectroscopy: In Vitro and In Oocytes Protein Structural Dynamics Studies,Angewandte Chemie International Edition

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A Bioresistant Nitroxide Spin Label for In‐Cell EPR Spectroscopy: In Vitro and In Oocytes Protein Structural Dynamics Studies
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2018-01-08 , DOI: 10.1002/anie.201710184
Ganesan Karthikeyan ₁ , Alessio Bonucci ₂ , Gilles Casano ₁ , Guillaume Gerbaud ₂ , Sébastien Abel ₁ , Virginie Thomé ₃ , Laurent Kodjabachian ₃ , Axel Magalon ₄ , Bruno Guigliarelli ₂ , Valérie Belle ₂ , Olivier Ouari ₁ , Elisabetta Mileo ₂

Affiliation

Approaching protein structural dynamics and protein–protein interactions in the cellular environment is a fundamental challenge. Owing to its absolute sensitivity and to its selectivity to paramagnetic species, site‐directed spin labeling (SDSL) combined with electron paramagnetic resonance (EPR) has the potential to evolve into an efficient method to follow conformational changes in proteins directly inside cells. Until now, the use of nitroxide‐based spin labels for in‐cell studies has represented a major hurdle because of their short persistence in the cellular context. The design and synthesis of the first maleimido‐proxyl‐based spin label (M‐TETPO) resistant towards reduction and being efficient to probe protein dynamics by continuous wave and pulsed EPR is presented. In particular, the extended lifetime of M‐TETPO enabled the study of structural features of a chaperone in the absence and presence of its binding partner at endogenous concentration directly inside cells.

中文翻译：

用于细胞内EPR光谱的生物抗性一氧化氮自旋标记：体外和卵母细胞中蛋白质结构动力学研究

在细胞环境中接近蛋白质结构动力学和蛋白质间相互作用是一项根本性的挑战。由于其绝对的敏感性和对顺磁性物质的选择性，定点自旋标记（SDSL）与电子顺磁共振（EPR）结合有可能发展成为一种有效的方法来直接追踪细胞内部蛋白质的构象变化。迄今为止，由于基于硝化氮的自旋标记物在细胞环境中的持续时间短，因此在细胞内研究中一直是一个主要障碍。介绍了第一个基于马来酰亚胺基脯氨酸的自旋标记（M-TETPO）的设计和合成，该标记具有抗还原作用，并且能够通过连续波和脉冲EPR有效探测蛋白质动力学。尤其是，

更新日期：2018-01-08

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