PMID: 12758083May 22, 2003Paper

Can Förster resonance energy transfer measurements uniquely position troponin residues on the actin filament? A case study in multiple-acceptor FRET

Journal of Molecular Biology
J M RobinsonHerbert C Cheung

Abstract

Straightforward interpretation of Förster resonance energy transfer (FRET) data in terms of the distance from donor-labeled troponon-tropomyosin (TnTm) to acceptor-labeled actin is complicated by the potential for energy transfer to acceptors on neighboring actin monomers (cross-transfer). Calculations indicate that cross-transfer can account for a substantial percentage of the total transfer efficiency. In some cases, this renders isolated FRET data uninterpretable. To overcome these limitations, we have developed an analysis method that incorporates cross-transfer and can, in principle, define the most probable (in the "least-squares" sense) position of a TnTm residue on the actin filament. The technique analyzes data from four or more FRET experiments using acceptors attached to different residues on actin. We have used this method to specify the coordinates of skeletal troponin I (sTnI) residue 133 relative to the actin filament under Mg(2+) and Ca(2+)-saturating conditions. Ca(2+)-activation causes the C terminus of the regulatory domain of TnI to move away from the actin surface by 6.3A, laterally along the actin surface toward actin subdomain 3 by 22.0A, and azimuthally toward the actin inner domain by 13.2A. This inform...Continue Reading

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Citations

Jun 24, 2004·Proceedings of the National Academy of Sciences of the United States of America·I L MedintzJ R Deschamps
Jun 18, 2004·Journal of Molecular Biology·John M RobinsonHerbert C Cheung
Feb 16, 2005·Journal of Muscle Research and Cell Motility·Monica X LiBrian D Sykes
Jul 30, 2003·Nature Structural Biology·Brian D Sykes
Nov 14, 2006·Nucleic Acids Research·Gil AlterovitzMarco F Ramoni
Sep 4, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Jeffrey F BreitMark N Gillespie
Sep 17, 2009·Journal of the American Chemical Society·Anthony HarrimanRaymond Ziessel
Aug 5, 2008·Biophysical Journal·John M RobinsonWenji Dong

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