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FRET analysis of actin/myosin association in aortic smooth muscle
Author(s) -
Black Jason Edward,
Wright Gary L.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.lb14-d
Subject(s) - myosin , förster resonance energy transfer , actin , contraction (grammar) , biophysics , chemistry , microfilament , muscle contraction , aorta , sarcomere , microbiology and biotechnology , anatomy , biology , medicine , myocyte , biochemistry , cytoskeleton , fluorescence , endocrinology , cell , physics , quantum mechanics
Recent evidence has indicated that contraction of A7r5 rat aorta smooth muscle cells is accompanied by a significant loss in actin stress fiber structure with dissociation of actin and myosin. Fluorescence resonance energy transfer (FRET) was used to compare the association of smooth muscle myosin with actin in rat aorta. FRET analysis of rat aortic rings at different intervals of contraction indicated significant increases in α‐actin/myosin at the initiation (79%) and plateau (67%) in force development but not during the intermediate period of slowly developing tension (−4%). By comparison, β‐actin/myosin complex was unchanged except during slow force development where the association was significantly decreased (−30%). Similar to α‐actin/myosin, Alexa 488‐phalloidin staining fluorescence indicated increased tissue F‐actin content at initiation (21%) and the plateau (62%) in force. FRET images indicated the development of thickened cables and patches of α‐actin/myosin in tissue throughout the interval of contraction. The results provide direct evidence of dynamic remodeling of the contractile protein during vascular smooth muscle contraction and suggest that FRET analysis may be a powerful tool for assessment of tissue protein‐protein associations. This work was supported by a Marshall Cardio science grant.