定点荧光标记(SDFL): TrIQ方法提供使用荧光探针Bimane的见解

IF 1.1 4区 物理与天体物理 Q4 PHYSICS, ATOMIC, MOLECULAR & CHEMICAL
Anthony D. Shumate, David L. Farrens
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引用次数: 0

摘要

这篇评论是为了纪念韦恩·哈贝尔教授的80岁生日,以及他对科学哲学和实践的许多贡献。他继续开拓使用位点定向自旋标记(SDSL)作为研究蛋白质结构和动力学的有力工具。受他的努力启发,我们的实验室一直致力于开发定点荧光标记(SDFL)作为SDSL的补充方法。在整个过程中,我们模仿了Hubbell博士和他的同事在开发SDSL过程中使用的许多新颖方法和概念。这里讨论了我们的一些方法,但不是以全面或详尽的方式,而是强调我们发现的SDFL方法是健壮的、可重复的,并且相对容易使用。具体而言,我们将重点介绍利用双烷荧光探针进行SDFL的开发和应用,以及如何利用双烷的独特性质来收集蛋白质结构和功能的信息。我们还回顾了过去我们如何使用双烷和SDFL来研究G蛋白偶联受体(gpcr)的激活及其与关键结合伙伴的相互作用的例子。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Site-Directed Fluorescence Labeling (SDFL): TrIQ Methods Provide Insights Using the Fluorescent Probe Bimane

Site-Directed Fluorescence Labeling (SDFL): TrIQ Methods Provide Insights Using the Fluorescent Probe Bimane

This review is in honor of Professor Wayne Hubbell’s 80th birthday, and his many contributions to the philosophy and practice of science. He continues to pioneer the use of site-directed spin labeling (SDSL) as a powerful tool for studying protein structure and dynamics. Inspired by his efforts, our lab has worked to develop site-directed fluorescence labeling (SDFL) as a complementary approach to SDSL. Throughout this process, we have emulated many of the novel methods and concepts Dr. Hubbell and his colleagues used during their development of SDSL. Some of our approaches are discussed here, not in a comprehensive or exhaustive way, but to highlight SDFL methods we have found to be robust, reproducible, and relatively easy to use. Specifically, we will focus on the development and application of SDFL using the fluorescent probe bimane, and how the unique properties of bimane can be exploited to glean information about protein structure and function. We also review past examples of how we have used bimane and SDFL to investigate the activation of G protein-coupled receptors (GPCRs) and their interaction with key binding partners.

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来源期刊
Applied Magnetic Resonance
Applied Magnetic Resonance 物理-光谱学
CiteScore
1.90
自引率
10.00%
发文量
59
审稿时长
2.3 months
期刊介绍: Applied Magnetic Resonance provides an international forum for the application of magnetic resonance in physics, chemistry, biology, medicine, geochemistry, ecology, engineering, and related fields. The contents include articles with a strong emphasis on new applications, and on new experimental methods. Additional features include book reviews and Letters to the Editor.
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