Key Points
- This study investigates the influence of phospholamban phosphorylation on Ca2+-dependent ATPase activity in cardiac sarcoplasmic reticulum.
- Transient kinetic analysis of ATPase dynamics via rapid quenching device after incubation with and without cyclic AMP-dependent protein kinase.
- Evaluation of calcium binding and EP formation in Ca2+-free and Ca2+-bound microsomes with varying ATP concentrations.
- Comparison of initial rates and maximal EP formation using phosphorylated versus non-phosphorylated preparations.
- In Ca2+-free microsomes, phosphorylated ATPase showed enhanced initial rates of EP formation and calcium binding compared to non-phosphorylated.
- Significant enhancement of EP formation occurred with greater than 10 µM ATP, while lower ATP concentrations reduced this effect.
- Phospholamban phosphorylation notably increases the rate of calcium translocation across the microsomal membrane.
Structured PICO
PPopulationCardiac microsomes (sarcoplasmic reticulum)
IInterventionIncubation with cAMP-dependent protein kinase (phosphorylation of phospholamban)
CComparatorIncubation without cAMP-dependent protein kinase
OOutcomeTransient kinetics of the formation of the phosphorylated intermediate EP of ATPase and calcium bindingsurrogate
Phosphorylation of phospholamban by cAMP-dependent protein kinase accelerates calcium binding and translocation across the cardiac sarcoplasmic reticulum membrane.