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  • Title: A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics.
    Author: Colombo J, Antkowiak A, Kogan K, Kotila T, Elliott J, Guillotin A, Lappalainen P, Michelot A.
    Journal: Nat Commun; 2021 Jan 22; 12(1):548. PubMed ID: 33483497.
    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.
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