Voltage-clamp fluorometry for advancing mechanistic understanding of ion channel mechanisms with a focus on acid-sensing ion channels.

IF 3.8 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemical Society transactions Pub Date : 2024-10-30 DOI:10.1042/BST20240165
Eleonora Centonze, Stephan Kellenberger
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Abstract

Voltage-clamp fluorometry (VCF) has revolutionized the study of ion channels by combining electrophysiology with fluorescence spectroscopy. VCF allows ion channel researchers to link dynamic structural changes, measured in real time, to function. Acid-sensing ion channels (ASICs) are Na+-permeable non-voltage-gated ion channels of the central and peripheral nervous system. They function as pH sensors, triggering neuronal excitation when pH decreases. Animal studies have shown the importance of ASICs for pain and fear sensation, learning, and neurodegeneration following ischaemic stroke. This review explores the technical bases and various developments of VCF, including fluorescence resonance energy transfer and the use of unnatural fluorescent amino acids. We provide an overview of VCF applications with a focus on ASICs, detailing how VCF has unveiled proton-induced conformational changes in key regions such as the acid pocket, wrist, and pore, crucial for understanding transitions between closed, open, and desensitized states.

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电压钳荧光测定法促进对离子通道机制的机理认识,重点关注酸感应离子通道。
电压钳荧光测定法(VCF)将电生理学与荧光光谱学相结合,彻底改变了离子通道研究。VCF 使离子通道研究人员能够将实时测量到的动态结构变化与功能联系起来。酸感应离子通道(ASIC)是中枢和外周神经系统中的Na+渗透性非电压门控离子通道。它们具有 pH 值传感器的功能,当 pH 值降低时会触发神经元兴奋。动物研究表明,ASIC 对缺血性中风后的疼痛和恐惧感觉、学习和神经变性具有重要作用。本综述探讨了 VCF 的技术基础和各种发展,包括荧光共振能量转移和非天然荧光氨基酸的使用。我们概述了 VCF 在 ASIC 上的应用,详细介绍了 VCF 如何揭示酸袋、腕和孔等关键区域中质子诱导的构象变化,这对于理解封闭、开放和脱敏状态之间的转变至关重要。
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来源期刊
Biochemical Society transactions
Biochemical Society transactions 生物-生化与分子生物学
CiteScore
7.80
自引率
0.00%
发文量
351
审稿时长
3-6 weeks
期刊介绍: Biochemical Society Transactions is the reviews journal of the Biochemical Society. Publishing concise reviews written by experts in the field, providing a timely snapshot of the latest developments across all areas of the molecular and cellular biosciences. Elevating our authors’ ideas and expertise, each review includes a perspectives section where authors offer comment on the latest advances, a glimpse of future challenges and highlighting the importance of associated research areas in far broader contexts.
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