Phosphoinositides in frog skeletal muscle: a quantitative analysis

The contents of major phospholipids per g of wet wt. in frog skeletal muscle are: 5.3 mumol PC; 1.4 mumol PE; 1 mumol SM; 0.4 mumol PtdIns; 0.3 mumol CL; and 0.13 mumol PS. The quantities of polyphosphoinositides per g of wet wt. are: 181 nmol PtInsP; 28 nmol PtdInsP2; and 8 nmol lyso-PtdInsP2. The specific activity of labelling of the total muscle ATP attained by external incubation with [32P]Pi was found to be 57 dpm/nmol x g muscle wet wt. PtdInsP2, the highest labelled polyphosphoinositide, showed a specific activity of 64,000 dpm/nmol per g muscle wet wt., suggesting that high specific activity ATP may be compartmentalized in the local environment of the triads and used as a substrate by the PtdIns and PtInsP kinase in that region. PtdInsP2 which is the immediate precursor for the release of InsP3, is found at a significant concentration and strategically located for its postulated role as a substrate for the action of phosphoinositidase C. The presence of a novel endogenous polyphosphoinositide, lyso-PtdInsP2, in animal tissues is reported for the first time. Electrical stimulation leads towards a rapid catabolization of polyphosphoinositides revealed by reductions in the 3H- and 32P-labelling, suggesting that muscle excitation is associated with the activation of breaking down of polyphosphoinositides.