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

Research output: Contribution to journalArticlepeer-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 languageEnglish
Pages (from-to)1013-1019
Number of pages7
JournalNature Methods
Volume9
Issue number10
DOIs
Publication statusPublished - Oct 2012

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Ultrafast force-clamp spectroscopy of single molecules reveals load dependence of myosin working stroke'. Together they form a unique fingerprint.

Cite this