Abstract
A technique has been developed in laboratory to estimate the extent to which Ca 2+ -calmodulin is bound to, and stimulates the activity of calmodulin-sensitive cyclic nucleotide phosphodiesterase in intact tissue. As the binding of calmodulin to the phosphodiesterase is influenced by changes in the Ca 2+ concentration between 0. 1 and 10μM, this technique might be used as a qualitative indicator of Ca 2+ concentration changes in the cellular compartment containing the target protein. This approach permits a single determination to be made on approximately 75 mg of frozen tissue and involves no exceptional treatment of the tissue, such as introduction of Ca 2+ indicator substances into the cell, prior to experimental treatment and subsequent freezing. This chapter describes this particular measurement for illustration. The usual application of this technique is to measure stimulation of calmodulin-sensitive cGMP phosphodiesterase activity in intact porcine coronary artery strips.
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