Evidence that phospholipase delta1 is the effector in the Gh (transglutaminase II)-mediated signaling.

A new class of GTP-binding protein transglutaminase II (Gh) couples to a 69-kDa phospholipase C (PLC). An 8-amino acid region (Leu665-Lys672) of the alpha-subunit of Gh (Galphah) is involved in interaction and activation of PLC, an observation that has now been used to characterize the 69-kDa PLC further. A 20-amino acid peptide corresponding to Leu654-Leu673 of Galphah was used to prepare an affinity resin. On incubation with a partially purified PLC preparation from rat liver membranes, the affinity resin-bound approximately69- and 85-kDa proteins were recognized by an antibody to the 69-kDa PLC. Both purified 69-kDa PLC and PLC-delta1 bound to the affinity resin; moreover, antibodies to PLC-delta1 recognized the 69-kDa PLC, and antibodies to the 69-kDa PLC recognized PLC-delta1. A synthetic peptide corresponding to Leu661-Lys672 of Galphah inhibited the binding of PLC-delta1 to the affinity resin and also stimulated PLC-delta1. Reconstitution of PLC-delta1 with GTPgammaS (guanosine 5'-3-O-(thio)triphosphate)-activated Gh resulted in activation of PLC-delta1. Antibodies to Galphah also coimmunoprecipitated PLC-delta1 upon activation of Gh. These findings indicate that PLC-delta1 is the effector of Gh-mediated signaling.