用于单分子荧光共振能量转移研究的选择性双标记无细胞合成蛋白质的制备
Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies.
作者信息
Sadoine Mayuri, Cerminara Michele, Fitter Jörg, Katranidis Alexandros
机构信息
Forschungszentrum Jülich, Institute of Complex Systems ICS-5, Jülich, Germany.
RWTH Aachen, I.Physikalisches Institut (IA), Aachen, Germany.
出版信息
Bio Protoc. 2018 Jun 20;8(12):e2881. doi: 10.21769/BioProtoc.2881.
Abstract
Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor fluorophores. Here, we present a detailed protocol for preparing double-labeled proteins for smFRET studies. The protocol describes two cell-free approaches to achieve a selective label scheme that allows the highest possible accuracy in inter-dye distance determination.
摘要
单分子荧光共振能量转移（smFRET）是研究生物分子的分子结构和构象变化的有力工具。该技术需要在特定位点配备一对供体和受体荧光团的蛋白质样品。在此，我们提供了一份用于制备用于smFRET研究的双标记蛋白质的详细方案。该方案描述了两种无细胞方法，以实现一种选择性标记方案，从而在染料间距离测定中达到尽可能高的准确性。
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本文引用的文献
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ACS Synth Biol. 2018 Feb 16;7(2):405-411. doi: 10.1021/acssynbio.7b00433. Epub 2018 Jan 31.
2
Selective Double-Labeling of Cell-Free Synthesized Proteins for More Accurate smFRET Studies.用于更准确 smFRET 研究的无细胞合成蛋白质的选择性双重标记。
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