Isolation of cardiac myofibrils and myosin light chains with in vivo levels of light chain phosphorylation - Holroyde, M.J.; Small, D.A.P.; Howe, E.; Solaro, R.J. (Elsevier)
Conditions are described for the preparation of functional myofibrils and myosin light chains from freeze-clamped beating hearts with the state of light chain phosphorylation chemically 'frozen' during the extraction procedure. Myofibrils were shown to be functionally intact by measurement of Ca^2^+ binding and ATPase activity. Highly purified cardiac myosin light chains could be routinely isolated from myofibrillar preparations using ethanol fractionation together with ion-exchange chromotography. Analysis of light chains for covalent phosphate indicated that basal levels of phosphorylation of the 18-20 000 dalton light chain of myosin in rabbit hearts beating in situ or in a perfusion apparatus were 0.3-0.4 mol/mol. Covalent phosphate content of the light chain fraction did not change during perfusion of hearts with 10 μM epinephrine.
http://linkinghub.elsevier.com/retrieve/pii/0304-4165(79)90014-X
Keywords: W; V; WW-YZ; U