Open AccessFraction of myosin cross-bridges bound to actin in active muscle fibers: estimation by fluorescence anisotropy measurements.
Author(s) -
Thomas P. Burghardt,
Katalin Ajtai
Publication year - 1985
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.82.24.8478
Subject(s) - fluorescence anisotropy , anisotropy , myosin , myosin head , actin , isometric exercise , perpendicular , fluorescence , biophysics , upper and lower bounds , chemistry , muscle contraction , molecular physics , physics , myosin light chain kinase , optics , anatomy , biology , geometry , biochemistry , physiology , mathematical analysis , mathematics
Time-resolved and steady-state fluorescence anisotropy measurements from fluorescence-labeled myosin cross-bridges in single glycerinated skeletal muscle fibers in rigor, relaxed, MgADP-induced, and contracting states have been made in order to estimate the fraction of actin-bound cross-bridges in active muscle. When the plane of polarization of the excitation light is perpendicular to the fiber axis and its propagation vector has a component parallel to this axis, actin-bound cross-bridge states, such as rigor and MgADP-induced, have time-zero and steady-state anisotropies that are substantially lower than has the relaxed state. This difference provides a means of determining the fraction of cross-bridges bound to actin in active isometric fibers, by comparing the fluorescence anisotropy from active fibers with the anisotropy from bound and unbound cross-bridges in static states. By assuming that the active cross-bridges are either bound (in the manner of rigor or MgADP-induced states) or relaxed, we estimate that greater than 80% of the cross-bridges are actin-bound in active isometric fibers.