Ultrafast force-clamp spectroscopy of single molecules reveals load dependence of myosin working stroke

Marco Capitanio; Monica Canepari; Manuela Maffei; Diego Beneventi; Carina Monico; Francesco Vanzi; Roberto Bottinelli; Francesco Saverio Pavone

doi:10.1038/nmeth.2152

Ultrafast force-clamp spectroscopy of single molecules reveals load dependence of myosin working stroke

Marco Capitanio, Monica Canepari, Manuela Maffei, Diego Beneventi, Carina Monico, Francesco Vanzi, Roberto Bottinelli, Francesco Saverio Pavone

Istituti Clinici Scientifici Maugeri Spa – Società Benefit

Research output: Contribution to journal › Article › peer-review

Abstract

We describe a dual-trap force-clamp configuration that applies constant loads between a binding protein and an intermittently interacting biological polymer. The method has a measurement delay of only ∼10 μs, allows detection of interactions as brief as ∼100 μs and probes sub-nanometer conformational changes with a time resolution of tens of microseconds. We tested our method on molecular motors and DNA-binding proteins. We could apply constant loads to a single motor domain of myosin before its working stroke was initiated (0.2-1 ms), thus directly measuring its load dependence. We found that, depending on the applied load, myosin weakly interacted (

Original language	English
Pages (from-to)	1013-1019
Number of pages	7
Journal	Nature Methods
Volume	9
Issue number	10
DOIs	https://doi.org/10.1038/nmeth.2152
Publication status	Published - Oct 2012

ASJC Scopus subject areas

Biotechnology
Molecular Biology
Biochemistry
Cell Biology

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10.1038/nmeth.2152

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AU - Capitanio, Marco

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AU - Maffei, Manuela

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AU - Monico, Carina

AU - Vanzi, Francesco

AU - Bottinelli, Roberto

AU - Pavone, Francesco Saverio

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Ultrafast force-clamp spectroscopy of single molecules reveals load dependence of myosin working stroke

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