PHOSPHOLIPASE-C ACTIVITY IN MEMBRANES AND A SOLUBLE FRACTION ISOLATEDFROM FROG SKELETAL-MUSCLE

Highly purified triads and transverse tubules, as well as a soluble fr action isolated from frog skeletal muscle, hydrolyze exogenous phospha tidylinositol 4,5-bisphosphate forming inositol 1,4,5-trisphosphate wi th maximal rates in the range 0.5-1 nmol/mg per min at pCa 3. Sarcopla smic reticulum membranes present a minor activity. The hydrolysis rate s in triads were 0.072 +/- 0.015 nmol/mg per min at pCa 7, increasing to 0.263 +/- 0.026 nmol/ mg per min at pCa 5 with 1.0 mM Mg and 0.1 mM substrate. The phospholipase C activity of isolated transverse tubule s at pCa 3 was 0.570 +/- 0.032 nmol/ mg per min. Since triads contain 10% transverse tubules, and correcting for the small contribution of s arcoplasmic reticulum, the calculated phospholipase C activity of tran sverse tubules at pCa 3 is about 10-times higher than the observed val ues, suggesting loss of activity during isolation. The activation by c alcium was also observed in a soluble fraction and was neither replace d nor inhibited by magnesium. No effect of GTP analogs on phospholipas e C activity was detected.