A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics

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Colombo , J , Antkowiak , A , Kogan , K , Kotila , T , Elliott , J , Guillotin , A , Lappalainen , P & Michelot , A 2021 , ' A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics ' , Nature Communications , vol. 12 , no. 1 , 548 . https://doi.org/10.1038/s41467-020-20827-4

Title: A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics
Author: Colombo, Jessica; Antkowiak, Adrien; Kogan, Konstantin; Kotila, Tommi; Elliott, Jenna; Guillotin, Audrey; Lappalainen, Pekka; Michelot, Alphée
Other contributor: University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
Date: 2021-01-22
Language: eng
Number of pages: 13
Belongs to series: Nature Communications
ISSN: 2041-1723
DOI: https://doi.org/10.1038/s41467-020-20827-4
URI: http://hdl.handle.net/10138/328018
Abstract: Actin polymerization provides force for vital processes of the eukaryotic cell, but our understanding of actin dynamics and energetics remains limited due to the lack of high-quality probes. Most current probes affect dynamics of actin or its interactions with actin-binding proteins (ABPs), and cannot track the bound nucleotide. Here, we identify a family of highly sensitive fluorescent nucleotide analogues structurally compatible with actin. We demonstrate that these fluorescent nucleotides bind to actin, maintain functional interactions with a number of essential ABPs, are hydrolyzed within actin filaments, and provide energy to power actin-based processes. These probes also enable monitoring actin assembly and nucleotide exchange with single-molecule microscopy and fluorescence anisotropy kinetics, therefore providing robust and highly versatile tools to study actin dynamics and functions of ABPs.
Subject: BINDING
COMPLEX
FILAMENT TURNOVER
NUCLEATION
POLYMERIZATION
PROFILIN
VISUALIZATION
1182 Biochemistry, cell and molecular biology
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