Subcellular distribution of phospholipid-sensitive calcium-dependent protein kinase in guinea pig heart, spleen and cerebral cortex, and inhibition of the enzyme by Triton X-100

Abstract

The subcellular distribution of phospholipid-sensitive Ca 2+-dependent protein kinase in guinea pig heart was found to be: cytosol, 73%; microsome, 18%; plasma membrane, 9%; nuclei and mitochondria, < 0.1%. The enzyme in spleen and cerebral cortex was distributed nearly equally in the cytosolic and total (unfractionated) particulate fractions. The particulate enzyme in heart was released by EGTA (2.5 mM) alone but not by Triton X-100 (0.3%) alone, although a combination of the two was most effective. On the other hand, the particulate enzyme in spleen and cerebral cortex was released only by a combination of Triton X-100 and EGTA. Triton X-100 inhibited the enzyme, and this inhibition was reversed by phosphatidylserine (a phospholipid cofactor for the enzyme). The detergent, however, was without effect on cyclic AMP-dependent and cyclic GMP-dependent protein kinases.