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

Jessica Colombo (1) , Adrien Antkowiak (1) , Konstantin Kogan (2) , Tommi Kotila (2) , Jenna Elliott (1) , Audrey Guillotin (1) , Pekka Lappalainen (2) , Alphee Michelot (1)

1 IBDM - Institut de Biologie du Développement de Marseille
2 Helsingin yliopisto = Helsingfors universitet = University of Helsinki

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 highquality probes. Most current probes affect dynamics of actin or its interactions with actinbinding 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.

Domaines: Biologie structurale [q-bio.BM]

Jessica Colombo, Adrien Antkowiak, Konstantin Kogan, Tommi Kotila, Jenna Elliott, et al.. A functional family of fluorescent nucleotide analogues to investigate actin dynamics and energetics. Nature Communications, 2021, 12, pp.548. ⟨10.1038/s41467-020-20827-4⟩. ⟨hal-03456460⟩