FRET 技术：利用基因编码荧光氨基酸进行距离依赖能量转移的案例研究。
FRETing about the details: Case studies in the use of a genetically encoded fluorescent amino acid for distance-dependent energy transfer.
机构信息
Graduate Group in Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA.
Department of Chemistry, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA.
出版信息
Protein Sci. 2023 May;32(5):e4633. doi: 10.1002/pro.4633.
Abstract
Förster resonance energy transfer (FRET) is a valuable method for monitoring protein conformation and biomolecular interactions. Intrinsically fluorescent amino acids that can be genetically encoded, such as acridonylalanine (Acd), are particularly useful for FRET studies. However, quantitative interpretation of FRET data to derive distance information requires careful use of controls and consideration of photophysical effects. Here we present two case studies illustrating how Acd can be used in FRET experiments to study small molecule induced conformational changes and multicomponent biomolecular complexes.
摘要
Förster 共振能量转移（FRET）是一种监测蛋白质构象和生物分子相互作用的有价值的方法。可以遗传编码的天然荧光氨基酸，如 acridonylalanine（Acd），特别适用于 FRET 研究。然而，为了从 FRET 数据中得出距离信息，需要仔细使用对照并考虑光物理效应。本文介绍了两个案例研究，说明了如何在 FRET 实验中使用 Acd 来研究小分子诱导的构象变化和多组分生物分子复合物。
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