基于超黄 pHluorin 的改进型基因编码 pH 传感器的合理设计,ACS Sensors

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基于超黄 pHluorin 的改进型基因编码 pH 传感器的合理设计
ACS Sensors ( IF 8.2 ) Pub Date : 2023-07-23 , DOI: 10.1021/acssensors.3c00484
Yi Shen ₁ , Yurong Wen _{2,

3} , Silvia Sposini _{4,

5} , Anjali Amrapali Vishwanath ₆ , Ahmed S Abdelfattah _{7,

8} , Eric R Schreiter ₇ , M Joanne Lemieux ₂ , Jaime de Juan-Sanz ₆ , David Perrais ₄ , Robert E Campbell _{1,

9}

Affiliation

Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
Department of Biochemistry, Faculty of Medicine & Dentistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
Center for Microbiome Research of Med-X Institute, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China.
CNRS, Interdisciplinary Institute for Neuroscience, UMR 5297, University of Bordeaux, Bordeaux 33076, France.
Department of Metabolism, Digestion and Reproduction, Institute of Reproductive and Developmental Biology, Imperial College London, London SW7 2BX, United Kingdom.
Institut du Cerveau-Paris Brain Institute-ICM, Inserm, CNRS, APHP, Häpital de la Pitié Salpêtrière, Sorbonne Université, 75013 Paris, France.
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virgina 20147, United States.
Department of Neuroscience, Brown University, Providence, Rhode Island 02906, United States.
Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan.

基于荧光蛋白的基因编码 pH 传感器是对与 pH 变化相关的细胞事件（例如胞吐作用和内吞作用）进行成像的宝贵工具。超黄 pHluorin (SEP) 是一种 pH 敏感的绿色荧光蛋白 (GFP) 变体，广泛用于此类应用。在此，我们报告了 Lime 的合理设计、开发、结构和应用，Lime 是一种改进的 SEP 变体，具有更高的荧光亮度和更高的 pH 敏感性。Lime 的 X 射线晶体结构支持了引导有益突变引入的机制原理。相对于 SEP，Lime 在胞吞作用和胞吐作用的成像方面提供了实质性改进。此外，Lime 及其变体有利于更广泛的应用，包括检测突触释放和神经元电压变化。

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更新日期：2023-07-23

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