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WITHDRAWN: Microsecond resolution of enzymatic conformational changes using dark-field microscopy

Authors :
Wayne D. Frasch
Justin York
James Martin
David Spetzler
Robert Ishmukhametov
Source :
Methods (San Diego, Calif.).
Publication Year :
2008

Abstract

We report a novel method to detect angular conformational changes of a molecular motor in a manner sensitive enough to achieve acquisition rates with a time resolution of 2.5mus (equivalent to 400,000fps). We show that this method has sufficient sensitivity to resolve the velocity of the F(1)-ATPase gamma-subunit as it travels from one conformational state to another (transition time). Rotation is detected via a gold nanorod attached to the rotating gamma-subunit of an immobilized F(1)-ATPase. Variations in scattered light intensity allow precise measurement of changes in angular position of the rod below the diffraction limit of light.

Details

Language :
English
ISSN :
10959130 and 10462023
Database :
OpenAIRE
Journal :
Methods (San Diego, Calif.)
Accession number :
edsair.doi.dedup.....f1adaecb1df2b7e2c5806828ac686550