Conventional isoforms of PKC are activated by the second messengers calcium and diacylglycerol (DAG). In the absence of these second messengers, PKC has a low membrane affinity and localizes primarily to the cytosol. Following agonist stimulation, calcium binds to the C2 domain (yellow), targeting it to anionic phospholipid membranes. PKC then undergoes an efficient 2-dimensional search for DAG in the plane of the membrane. Membrane-partitioned DAG and phosphatidylserine (PS) help anchor PKC to membranes by binding the C1 domain (orange). When both domains are engaged, an autoinhibitory pseudosubstrate sequence (purple) is expelled from the active site of the catalytic domain (blue). The exposed active site catalyzes the transfer of phosphate to suitably localized protein substrates. As cells return to the resting state, levels of calcium and DAG are reduced, inactivating PKC by lowering its membrane affinity.