Tension Transients in Skinned Muscle Fibres of Insect Flight Muscle and Mammalian Cardiac Muscle: Effect of Substrate Concentration and Treatment with Myosin Light Chain Kinase
Glycerinated single fibres from the dorsal longitudinal muscle of Lethocerus maximus were isometrically contracted in MgATP-salines (10 μM Ca2+; 1.5 mM Mg2+; pH 6.7: 22°C and 20 mM PEP; 100 U/ml pyruvate kinase). The ratio of ATPase activity to tension decreased by a factor of 2 after reducing the ATP-concentration from 15 to 0.5 mM. At all ATP-concentrations (0.5 – 15 mM), the fibres showed tension adjustments in response to small step changes in length characteristic to an actively contracting muscle: i) an elastic phase which did not depend on ATP-concentration ii) a quick phase of stress relaxation with at least two exponential components: iii) a phase of delayed tension generation. An increase in size of the length step and/or a decrease of ATP-concentration slowed the quick phase and the delayed phase Similar results have been obtained with skinned cardiac muscle (pig right ventricle). To see, how the isolated contractile system is affected by an increase in the light chain phosphorylation, tension transients were studied in skinned right ventricular muscle fibres before and after incubation with ATPγS (2 mM), pure myosin light chain kinase (9 μg/ml), Calmodulin (1 μM) and Cat2+ (0.8 μM). While isometric tension development elicited by 20 μM Ca2+ in the ATP salt solution was barely affected in presence of the enzyme, the ATPase activity was decreased by about 25% of the control. There was also a marked decrease (about 50%) in the contraction velocity as determined by the recovery of tension following a quick release. Quick stretches cause an immediate increase in tension followed by a rapid fall and a subsequent rise in tension. The velocity of this tension rise decreased by approximately 30% after incubation with myosin light chain kinase.
KeywordsATPase Activity Myosin Light Chain Kinase Myosin Light Chain Phosphorylation Quick Release Quick Phase
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