Inhibition of pancreatic phospholipase A2 activity by uteroglobin and antiflammin peptides: possible mechanism of action.
We investigated the possible mechanism of inhibition of porcine pancreatic phospholipase A2 in vitro by rabbit uteroglobin and by the antiflammin peptides. We optimized the conditions of phospholipase A2 assay using a deoxycholate-phosphatidylcholine mixed micellar substrate and established the activity of these inhibitors under optimized conditions. The results of fluorescence studies and crosslinking experiments indicate that the inhibitors interact with the enzyme in solution and affect the increase in intrinsic fluorescence of phospholipase A2 observed upon interaction with a mixed micellar substrate. In addition, we identified a sequence similarity between the antiflammin peptides, the putative active region of uteroglobin and a region in pancreatic phospholipase A2. This region of phospholipase A2 has been previously identified as being involved in the regulation of dimerization of this enzyme, and is conserved in the pancreatic-type enzymes. Taken together, these observations suggest that uteroglobin and antiflammins interact with porcine pancreatic phospholipase A2 and this may, at least in part, explain the enzyme inhibitory effect of these molecules observed in vitro. One possible mechanism of this effect may be an interference with the dimerization process of phospholipase A2 which is associated with interfacial activation.
Facchiano
A
A
Section on Developmental Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Cordella-Miele
E
E
Miele
L
L
Mukherjee
A B
AB
D000595:Amino Acid Sequence; D000818:Animals; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000494:pharmacology*; D010179:Pancreas / Q000201:enzymology*; D010446:Peptide Fragments / Q000494:pharmacology*; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000378:metabolism; D054467:Phospholipases A2; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D014598:Uteroglobin / Q000494:pharmacology*
D003432:Cross-Linking Reagents; D009842:Oligopeptides; D010446:Peptide Fragments; C064645:antiflammin P1; C064647:antiflammin P2; D014598:Uteroglobin; D010741:Phospholipases A; D054467:Phospholipases A2