Hydrolysis of p‐nitrophenyl acetate by the peptide chain fragment (336–449) of porcine pancreatic lipase
- 1 August 1986
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 158 (3), 601-607
- https://doi.org/10.1111/j.1432-1033.1986.tb09797.x
Abstract
The incubation of porcine pancreatic lipase (449 amino acids) with chymotrypsin led to the preferential cleavage of the Phe-335-Ala-336 bond [Bousset-Risso et al. (1985) FEBS Lett. 182, 323-326]. Up to now it has not been possible to isolate the fragment (1-335) whereas fragment (336-449) was purified. This fragment does not display any activity towards the specific substrates of lipase, triacylglycerols, either in the aggregate form or monomeric solution, but like lipase it hydrolyzes p-nitrophenyl acetate. The biphasic kinetics of the release of p-nitrophenol by the fragment with different concentrations of p-nitrophenyl acetate ([S] greater than [E]) are very similar to those of lipase and other esterases. The initial burst is equal to 1 mol p-nitrophenol/mol fragment (when [S] = infinity). Ethoxyformic anhydride only reacts with 1 mol histidine out of the 2 mol that the fragment contained. The activity of the fragment towards p-nitrophenyl acetate hydrolysis is inhibited after ethoxyformic anhydride reaction as in the case of lipase. The results presented led to the hypothesis that in the area (336-449) a part of the active-site structure of the lipase molecule is included. It would seem that fragment (336-449) is a functional domain of lipase.Keywords
This publication has 20 references indexed in Scilit:
- Limited proteolysis of porcine pancreatic lipaseFEBS Letters, 1985
- Porcine pancreatic lipase. Completion of the primary structureBiochimica et Biophysica Acta (BBA) - Protein Structure, 1981
- Direct determination of the protonation states of aspartic acid-102 and histidine-57 in the tetrahedral intermediate of the serine proteases: neutron structure of trypsinBiochemistry, 1981
- The reactive tyrosine residue of human serum albumin: Characterization of its reaction with diisopropylfluorophosphateArchives of Biochemistry and Biophysics, 1979
- An improved large scale procedure for the purification of porcine pancreatic lipaseBiochimica et Biophysica Acta (BBA) - Enzymology, 1978
- Mechanism of pancreatic lipase action. 2. Catalytic properties of modified lipasesBiochemistry, 1976
- Acetylation of human serum albumin by p-nitrophenyl acetateBiochemistry, 1975
- On the transient formation of an acetyl enzyme intermediate during the hydrolysis of P-nitrophenyl acetate by pancreatic lipaseBiochemical and Biophysical Research Communications, 1974
- Action of pancreatic lipase on monomeric tripropionin in the presence of water-miscible organic compoundsBiochimica et Biophysica Acta (BBA) - Enzymology, 1974
- The Kinetics of the α-Chymotrypsin-Catalyzed Hydrolysis of p-Nitrophenyl Acetate*Biochemistry, 1962