Excessive production of eicosanoids is characteristic of many inflammatory diseases. In this study we show that ceramide, which is an early messenger of inflammatory cytokine action, exerts a dual effect on the cytosolic phospholipase A₂ (cPLA₂), the rate‐limiting enzyme in arachidonic acid release and subsequent eicosanoid formation. Stimulation of renal mesangial cells with exogenous short‐chain ceramide analogs for 30 and 60 min leads to a concentration‐dependent increase in arachidonic acid release that is not blocked by specific inhibitors of mitogen‐activated protein kinase pathways. This suggests that these established upstream activators of cPLA₂ are not involved in ceramide‐induced arachidonic acid release. By use of photoactivatable ceramide analogs, D‐ and L‐ [¹²⁵I]3‐trifluoromethyl‐3‐(m‐iodophenyl)diazirineceramides (TID‐ceramides), we observed a direct interaction of ceramide with cPLA₂. This interaction was independent of the absolute configuration as D‐ and L‐TID‐ceramide were equally effective in binding to cPLA₂. Moreover, recombinant CaLB domain of cPLA₂ as well as a mutant deficient in the connecting ‘hinge’ domain of cPLA₂, efficiently bound D‐ and L‐TID‐ceramides, whereas the catalytic domain did not interact with TID‐ceramides. In vitro binding assays reveal that stearoyl‐arachidonyl‐phosphatidylcholine (SAPC)‐liposomes containing increasing mol% of ceramide lead to an increased association of recombinant cPLA₂ to the liposomes. Furthermore, measurement of cPLA₂ activity in vitro shows that the presence of SAPC‐liposomes resulted in only weak cPLA₂ activity. However, the activity dramatically increases by addition of ceramide to the liposomes. Furthermore, liposomes containing SAPC and sphingomyelin resulted in no better substrate than SAPC liposomes, unless bacterial sphingomyelinase was added to generate ceramide, which then causes a marked increase in cPLA₂ activity. These results demonstrate that ceramide can interact directly with cPLA₂ via the CaLB domain and thereby serves as a membrane‐docking device that facilitates cPLA₂ action in inflammatory diseases.