Substantial amounts of platelet-activating factor (PAF 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine), the potent phospholipid mediator of allergic and inflammatory reactions, are formed upon incubation of acetylated low-density lipoprotein, acetylated bovine serum albumin (BSA) and acetylated apolipoprotein A-I with 1-0-hexadecyl-sn-glycero-3-phosphocholine (lyso-PAF). Acetylated BSA produced 0.3 nmol PAF/mg of protein after a 6 h incubation period with 40 lM lyso-PAF. The transfer of acetate bound to acetylated proteins to lyso-PAF was non-enzymatic. Chemical PAF formation by acetylated proteins, involved in lipid metabolism and transport, could lead to complication of inflammatory and allergic events.