Characterization of acylating and deacylating activities of an extracellular phospholipase A2 in a water-restricted environment.
The behavior of porcine pancreatic phospholipase A2 (ppPLA2) in monophasic low-water media has been explored, for the first time, in a systematic manner. It has been investigated how a number of variables can modulate both acylating and deacylating activities of the enzyme, and several interesting, unexpected results are presented. Among the most relevant, when placing ppPLA2 in the water-restricted environment, are the following: (i) it displays a remarkable alteration of its specificity toward the substrate polar head relative to all-water medium; (ii) it is quite severely inhibited by lysophosphatidylcholine (LPC), which has important implications, particularly concerning its acylation activity; and (iii) it exquisitely discriminates between saturated and unsaturated long-chain fatty acids when esterifying them with LPC. Finally, it is also illustrated how these results can be exploited to optimize the catalytic performance of the enzyme in nonaqueous medium and obtain a nearly 30-fold increase in the yield of phosphatidylcholine synthesis with respect to previously reported data.
Mingarro
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Departament de Bioquímica i Biologia Molecular, Universitat de València, Burjassot, Spain.
Abad
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Braco
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D000215:Acylation; D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004795:Enzyme Stability; D004951:Esterification; D005110:Extracellular Space / Q000201:enzymology; D006358:Hot Temperature; D006868:Hydrolysis; D007700:Kinetics; D008244:Lysophosphatidylcholines / Q000378:metabolism; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology; D010713:Phosphatidylcholines / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D013379:Substrate Specificity; D013552:Swine; D014867:Water
D008244:Lysophosphatidylcholines; D009829:Oleic Acids; D010713:Phosphatidylcholines; D014867:Water; D019301:Oleic Acid; D004492:Edetic Acid; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium