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Proceedings Paper

Analysis of single-molecule mechanical measurements with high spatio-temporal resolution
Author(s): Marco Capitanio; Lucia Gardini; Francesco Saverio Pavone
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Paper Abstract

Optical tweezers allow recording mechanical data from single biological molecules such as molecular motors, DNA processing enzymes, nucleic acids. Such data consist of time series that are dominated by thermal noise and such noisy recordings require proper analysis to correctly extract kinetic and mechanical information. Several different analysis approaches have been established in the past years. Here, we propose an analysis method for optical trapping recordings of non-processive motor proteins. The method does not assume any particular interaction kinetics, allows detection of sub-millisecond interactions and quantification of the number of false and lost events. Precise alignment of interaction events and ensemble averaging allow the investigation of the stepping dynamics of non-processive motors with a temporal resolution of few tens of microseconds and a spatial resolution of few angstroms. Our analysis is applied to the study of the motor protein myosin from fast skeletal muscle. Thanks to the high spatio-temporal resolution, we can distinguish three mechanical pathways in the acto-myosin interaction, with several orders of magnitude different kinetics, which contribute in a load-dependent manner to the myosin working stroke.

Paper Details

Date Published: 12 September 2013
PDF: 6 pages
Proc. SPIE 8810, Optical Trapping and Optical Micromanipulation X, 881034 (12 September 2013); doi: 10.1117/12.2030455
Show Author Affiliations
Marco Capitanio, European Lab. for Non-linear Spectroscopy (Italy)
Univ. degli Studi di Firenze (Italy)
Lucia Gardini, European Lab. for Non-linear Spectroscopy (Italy)
Francesco Saverio Pavone, European Lab. for Non-linear Spectroscopy (Italy)
Univ. degli Studi di Firenze (Italy)
National Institute of Optics-CNR and International Ctr. of Computational Neurophotonics (Italy)

Published in SPIE Proceedings Vol. 8810:
Optical Trapping and Optical Micromanipulation X
Kishan Dholakia; Gabriel C. Spalding, Editor(s)

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