用光控制工程化P2X受体。

Controlling Engineered P2X Receptors with Light.

作者信息

Atkinson Benjamin N, Chudasama Vijay, Browne Liam E

机构信息

Department of Chemistry, University College London, London, UK.

Wolfson Institute for Biomedical Research, University College London, London, UK.

出版信息

Methods Mol Biol. 2020;2041:301-309. doi: 10.1007/978-1-4939-9717-6_22.

DOI:10.1007/978-1-4939-9717-6_22

PMID:31646498

Abstract

This chapter details methods to express and modify ATP-gated P2X receptor channels so that they can be controlled using light. Following expression in cells, a photoswitchable tool compound can be used to covalently modify mutant P2X receptors, as previously demonstrated for homomeric P2X2 and P2X3 receptors, and heteromeric P2X2/3 receptors. Engineered P2X receptors can be rapidly and reversibly opened and closed by different wavelengths of light. Light-activated P2X receptors can be mutated further to impart ATP-insensitivity if required. This method offers control of specific P2X receptor channels with high spatiotemporal precision to study their roles in physiology and pathophysiology.

摘要

本章详细介绍了表达和修饰三磷酸腺苷门控P2X受体通道的方法，以便可以用光对其进行控制。如之前在同聚体P2X2和P2X3受体以及异聚体P2X2/3受体中所证明的那样，在细胞中表达后，一种可光开关的工具化合物可用于共价修饰突变型P2X受体。工程化的P2X受体可以通过不同波长的光快速且可逆地打开和关闭。如果需要，光激活的P2X受体可以进一步突变以赋予对三磷酸腺苷不敏感的特性。该方法能够以高时空精度控制特定的P2X受体通道，以研究它们在生理和病理生理中的作用。