Induction of intracellular signalling by cyclic glycerophosphates and their deoxy analogues.

Abstract

Cyclic glycerophosphates can be formed by enzymic degradation of phospholipids. They have only recently attracted attention, and their physiological function is still obscure. In this study, we have searched for signalling functions of the natural 1,3-cyclic and 1,2-cyclic glycerophosphates, their deoxy analogues, and the phenyl esters of the 1,3-cyclic phosphates. Linear sn-glycerol 3-phosphate and glycerol 2-phosphate served as the control compounds. Each of the six-membered ring cyclic phosphates tested induced rapid intracellular tyrosine phosphorylation in CHO and NIH-3T3 cells when applied extracellularly at a concentration of 0.5-4 microM. The phosphorylated intracellular proteins had molecular masses of approximately 35 kDa, approximately 45 kDa, 60-70 kDa and approximately 120 kDa. The five-membered ring cyclic phosphates had a similar effect, but at an external concentration of 2-10 microM, while sn-glycerol 3-phosphate and glycerol 2-phosphate had no effect. The six-membered cyclic phosphates also induced rapid threonine phosphorylation in CHO cells of approximately 18-kDa, approximately 35-kDa, and approximately 38-kDa proteins. Further experiments indicated that the cyclic phosphates partition rapidly into the cell cytosol where they activate kinases, including mitogen-activated protein kinase. When their intracellular level increases, dephosphorylation presumably takes place. This pattern may account for the signalling profile of cyclic phosphates and suggests that they may take part in processes associated with cell differentiation.