1e37
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PORCINE PANCREATIC ELASTASE COMPLEXED WITH (3S, 4S)N-PARA-NITROBENZENESULPHONYL-3-ETHYL-4-(CARBOXYLIC ACID)PYRROLIDIN-2-ONE SOAKED IN PH 9 BUFFER FOR 1 MINUTE
OverviewOverview
beta-Lactams inhibit a range of enzymes via acylation of nucleophilic, serine residues. Certain gamma-lactam analogues of monocyclic beta-lactams, have also been shown to be reversible inhibitors of porcine pancreatic, elastase (PPE), forming acyl-enzyme complexes that are stable with respect, to hydrolysis. Crystallographic analysis at pH 5 of an acyl-enzyme complex, formed with PPE and one of these inhibitors revealed the ester carbonyl, located in the oxyanion hole in a similar conformation to that observed in, the structure of a complex formed between a heptapeptide, (beta-casomorphin-7) and PPE. Only weak electron density was observed for, the His-57 side chain in its 'native' conformation. Instead, the His-57, side chain predominantly adopted a conformation rotated approx. 90 degrees, from its normal position. PPE-gamma-lactam crystals were subjected to, 'pH-jumps' by placing the crystals in a buffer of increased pH prior to, freezing for data collection. The results indicate that the conformation, of the gamma-lactam-derived acyl-enzyme species in the PPE active site is, dependent on pH, a result having implications for the analysis of other, serine protease-inhibitor structures at non-catalytic pH values. The, results help to define the stereoelectronic relationship between the ester, of the acyl-enzyme complex, the side chain of His-57 and the incoming, nucleophile during the reversible (de)acylation steps, implying it is, closely analogous to the hydrolytic deacylation step during catalytic, peptide hydrolysis.
About this StructureAbout this Structure
1E37 is a Single protein structure of sequence from Sus scrofa with , and as ligands. Active as Pancreatic elastase, with EC number 3.4.21.36 Known structural/functional Site: . Full crystallographic information is available from OCA.
ReferenceReference
'pH-jump' crystallographic analyses of gamma-lactam-porcine pancreatic elastase complexes., Wright PA, Wilmouth RC, Clifton IJ, Schofield CJ, Biochem J. 2000 Oct 15;351 Pt 2:335-40. PMID:11023818
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