Background:The mechanistic regulation of phagocytosis by phospholipase A 2 (PLA 2 ) remains to be defined. Results: A specific PLA 2 isoform is activated during phagocytosis, translocates to the plasma membrane, and directly participates in phagosome formation, without involving its enzymatic activity. Conclusion: PLA 2 IV␣ regulates phagocytosis via a novel mechanism that requires membrane binding of its C2 domain. Significance: PLA 2 IV␣ mediates a novel mechanism of phagocytosis regulation.