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An integrated in vitro and in situ study of kinetics of myosin II from frog skeletal muscle
Author(s) -
Elangovan R.,
Capitanio M.,
Melli L.,
Pavone F. S.,
Lombardi V.,
Piazzesi G.
Publication year - 2012
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2011.222984
Subject(s) - myosin , sarcomere , actin , skeletal muscle , biophysics , myosin atpase , meromyosin , in situ , chemistry , myosin head , biology , myocyte , atpase , anatomy , microbiology and biotechnology , biochemistry , myosin light chain kinase , enzyme , organic chemistry
Key points • Force and shortening in muscle are due to the ATP‐powered motor protein myosin II, polymerized in two bipolar arrays of motors that pull the two overlapping actin filaments toward the centre of the sarcomere. • The parameters of the myosin motor in situ have been best characterized for the skeletal muscle of the frog, from which single intact cells can be isolated allowing fast sarcomere level mechanics to be applied. • Up to now no reliable methods have been developed for the study of frog myosin with single molecule techniques. • In this work a new protocol for extraction and conservation of frog muscle myosin allows us to estimate the sliding velocity of actin on myosin ( V F ) and its modulation by pH, myosin density, temperature and substrate concentration. • By integrating in vitro and in situ parameters of frog muscle myosin we can relate kinetic and mechanical steps of the acto‐myosin ATPase.