Analysis of single-molecule mechanical measurements with high spatio-temporal resolution

Marco Capitanio; Lucia Gardini; Francesco Saverio Pavone

doi:10.1117/12.2030455

12 September 2013 Analysis of single-molecule mechanical measurements with high spatio-temporal resolution

Marco Capitanio, Lucia Gardini, Francesco Saverio Pavone

Proceedings Volume 8810, Optical Trapping and Optical Micromanipulation X; 881034 (2013) https://doi.org/10.1117/12.2030455
Event: SPIE NanoScience + Engineering, 2013, San Diego, California, United States

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.

Citation Download Citation

Marco Capitanio, Lucia Gardini, and Francesco Saverio Pavone "Analysis of single-molecule mechanical measurements with high spatio-temporal resolution", Proc. SPIE 8810, Optical Trapping and Optical Micromanipulation X, 881034 (12 September 2013); https://doi.org/10.1117/12.2030455

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