While the role of the group IVA Ca²⁺-dependent cytosolic phospholipase A₂α (cPLA₂α) in arachidonic acid (AA) metabolism has been well documented, that of its paralogue, Ca²⁺-independent group IVC PLA₂ (cPLA₂γ), has remained uncertain. Here we show, using a transfection strategy, that cPLA₂γ has the ability to increase the spontaneous and stimulus-induced release of cellular fatty acids. The AA released by cPLA₂γ was metabolized further to prostaglandin E₂ via cyclo-oxygenase-1 (COX-1) in the immediate response, and via COX-2 in the delayed response. Mutation of the putative catalytic-centre residue Ser⁸² abrogated the AA-releasing function of cPLA₂γ both in vitro and in vivo. Confocal microscopy revealed that cPLA₂γ was distributed in the perinuclear endoplasmic reticulum membranes. Mutating the C-terminal prenylation site of cPLA₂γ abrogated its intracellular membrane localization and cellular AA-releasing function, without reducing its enzyme activity in vitro. Our results indicate that cPLA₂γ is the second cPLA₂ enzyme that contributes to cellular AA metabolism and phospholipid remodelling under appropriate conditions.