Competitive inhibition of lipolytic enzymes. III. Some acylamino analogues of phospholipids are potent competitive inhibitors of porcine pancreatic phospholipase A2.
Competitive inhibition of porcine pancreatic phospholipase A2 was studied in mixed-micellar systems composed of long- and medium-chain substrates, potential inhibitors and detergents. A number of positional and stereoisomeric monoacylamino, acyloxyglycerophospholipids were investigated for their inhibitory properties, using as substrates the corresponding diacyl-sn-glycero-3-phospholipids possessing the same polar headgroup and identical acyl chain lengths. Based on a kinetic model applicable to water-insoluble inhibitors (see accompanying paper I), which allows a quantitative comparison of the inhibitory power (Z) of the various phospholipid analogues, the following results were obtained: Substitution of a single acylester bond in a diacylglycerophospholipid by an acylamino group can transform the substrate molecule in a potent competitive inhibitor. This property is acquired only when this substitution occurs on the phospholipase-susceptible ester bond of the substrate. If the acylamino group replaces an ester bond which cannot be attacked by the highly positional and stereospecific phospholipase, the resulting molecule binds with similar affinity to the active site of the enzyme as the parent substrate molecule. Because of its positional and stereospecificity, this so-called inhibitory 'amide effect' suggests that these inhibitors behave as substrate-derived analogues. The inhibitory 'amide-effect' observed with several medium- and long-chain monoacyloxy-, monoacylamino-deoxyglycerophosphatides is completely lost upon specific alkaline hydrolysis of the single acylester bond. Reesterification of the free glycerol OH group in these lysoacylaminophosphoglycerides, even with an acetyl residue, restores the inhibitory properties. These observations indicate that specific binding of phospholipids to the active site of pancreatic phospholipase A2, requires the presence of two chains in substrate or inhibitor structure and suggest that those results obtained with lysophospholipids and single-chain analogues may be questionable.
de Haas
G H
GH
Laboratory of Biochemistry, C.B.L.E., Utrecht, The Netherlands.
Dijkman
R
R
van Oort
M G
MG
Verger
R
R
D000818:Animals; D001667:Binding, Competitive; D004791:Enzyme Inhibitors; D007700:Kinetics; D008066:Lipolysis; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D015394:Molecular Structure; D010179:Pancreas / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000494:pharmacology*; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D010742:Phospholipid Ethers / Q000378:metabolism; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013552:Swine
D004791:Enzyme Inhibitors; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D010742:Phospholipid Ethers; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2