Quantifying Instrumental Artifacts in Folding Kinetics Measured by Single-Molecule Force Spectroscopy.

Abstract:

:Force spectroscopy is commonly used to measure the kinetics of processes occurring in single biological molecules. These measurements involve attaching the molecule of interest to micron-sized or larger force probes via compliant linkers. Recent theoretical work has described how the properties of the probes and linkers can alter the observed kinetics from the intrinsic behavior of the molecule in isolation. We applied this theory to estimate the errors in measurements of folding made using optical tweezers. Errors in the folding rates arising from instrument artifacts were only ∼20% for constant-force measurements of DNA hairpins with typical choices of linker length and probe size. Measurements of transition paths using a constant trap position at high trap stiffness were also found to be in the low-artifact limit. These results indicate that typical optical trap measurements of kinetics reflect the dynamics of the molecule fairly well, and suggest practical limitations on experimental design to ensure reliable kinetic measurements.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Neupane K,Woodside MT

doi

10.1016/j.bpj.2016.06.011

subject

Has Abstract

pub_date

2016-07-26 00:00:00

pages

283-286

issue

2

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(16)30449-0

journal_volume

111

pub_type

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