Tropomyosin Dynamics in Cardiac Thin Filaments: A Multisite Förster Resonance Energy Transfer and Anisotropy Study

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dc.contributor.authorWang, Huien_US
dc.contributor.authorMao, Shuen_US
dc.contributor.authorChalovich, Josephen_US
dc.contributor.authorMarriott, Gerarden_US
dc.date.accessioned2011-01-28T19:37:41Zen_US
dc.date.accessioned2011-05-17T01:27:01Z
dc.date.available2011-01-28T19:37:41Zen_US
dc.date.available2011-05-17T01:27:01Z
dc.date.issued2008-06en_US
dc.description.abstractCryoelectron microscopy studies have identified distinct locations of tropomyosin (Tm) within the Ca21-free, Ca21-saturated, and myosin-S1-saturated states of the thin filament. On the other hand, steady-state Förster resonance energy transfer (FRET) studies using functional, reconstituted thin filaments under physiological conditions of temperature and solvent have failed to detect any movement of Tm upon Ca21 binding. In this investigation, an optimized system for FRET and anisotropy analyses of cardiac tropomyosin (cTm) dynamics was developed that employed a single tethered donor probe within a Tm dimer. Multisite FRET and fluorescence anisotropy analyses showed that S1 binding to Ca21 thin filaments triggered a uniform displacement of cTm toward F-actin but that Ca21 binding alone did not change FRET efficiency, most likely due to thermally driven fluctuations of cTm on the thin filament that decreased the effective separation of the donor probe between the blocked and closed states. Although Ca21 binding to the thin filament did not significantly change FRET efficiency, such a change was demonstrated when the thin filament was partially saturated with S1. FRET was also used to show that stoichiometric binding of S1 to Ca21-activated thin filaments decreased the amplitude of Tm fluctuations and revealed a strong correlation between the cooperative binding of S1 to the closed state and the movement of cTm. Originally published Biophysical Journal, Vol. 94, No. 11, June 2008en_US
dc.identifier.citationBiophysical Journal; 94:11 p. 4358-4369en_US
dc.identifier.pmidPMC2480674en_US
dc.identifier.urihttp://hdl.handle.net/10342/3133en_US
dc.language.isoen_USen_US
dc.publisherEast Carolina Universityen_US
dc.relation.urihttp://www.cell.com/biophysj/archiveen_US
dc.rightsAuthor notified of opt-out rights by Cammie Jenningsen_US
dc.subjectCryoelectron microscopyen_US
dc.subjectCardiac thin filamentsen_US
dc.subjectTropomyosinen_US
dc.titleTropomyosin Dynamics in Cardiac Thin Filaments: A Multisite Förster Resonance Energy Transfer and Anisotropy Studyen_US
dc.typeArticleen_US

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