Crystallographic analysis of the inhibition of porcine pancreatic elastase by a peptidyl boronic acid: structure of a reaction intermediate

Biochemistry ◽  
1989 ◽  
Vol 28 (19) ◽  
pp. 7610-7617 ◽  
Author(s):  
Lori H. Takahashi ◽  
R. Radhakrishnan ◽  
Richard E. Rosenfield ◽  
Edgar F. Meyer
Keyword(s):  
Boronic Acid ◽  
Crystallographic Analysis ◽  
Pancreatic Elastase ◽  
Reaction Intermediate ◽  
Porcine Pancreatic Elastase ◽  
Acid Structure

scholarly journals ‘pH-jump’ crystallographic analyses of γ-lactam–porcine pancreatic elastase complexes

2000 ◽  
Vol 351 (2) ◽  
pp. 335-340
Author(s):  
Penny A. WRIGHT ◽  
Rupert C. WILMOUTH ◽  
Ian J. CLIFTON ◽  
Christopher J. SCHOFIELD
Keyword(s):  
Crystallographic Analysis ◽  
Side Chain ◽  
Enzyme Complex ◽  
Peptide Hydrolysis ◽  
Reversible Inhibitors ◽  
Pancreatic Elastase ◽  
Ph Jump ◽  
Porcine Pancreatic Elastase ◽  
Enzyme Complexes ◽  
Similar Conformation

β-Lactams inhibit a range of enzymes via acylation of nucleophilic serine residues. Certain γ-lactam analogues of monocyclic β-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 (β-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° from its normal position. PPE–γ-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 γ-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.

The Effects of Various Lipids upon the Rate of Elastolysis of Bovine Ligamentum Nuchae Elastin by Porcine Pancreatic Elastase

1985 ◽  
Vol 68 (s11) ◽  
pp. 26P-27P
Author(s):  
S A Wharton ◽  
G J Phillips ◽  
Jmdc Pereira ◽  
Dcs Hutchison ◽  
H Baum
Keyword(s):  
Pancreatic Elastase ◽  
Porcine Pancreatic Elastase

scholarly journals Risk Assessment for Use of a Porcine Circovirus-Contaminated Reagent in a Barrier Maintained Rodent Colony

2020 ◽  
Vol 59 (5) ◽  
pp. 575-579
Author(s):  
Norman C Peterson ◽  
Aaron A Berlin
Keyword(s):  
Risk Assessment ◽  
Mouse Model ◽  
Cell Cultures ◽  
Direct Contact ◽  
Model Development ◽  
Porcine Circovirus ◽  
Laboratory Mice ◽  
Wild Mice ◽  
Pancreatic Elastase ◽  
Porcine Pancreatic Elastase

A proposal for the use of porcine pancreatic elastase (PPE) to develop a mouse model of pulmonary emphysema raised concerns about introducing contaminating porcine viruses into our barrier facility. Porcine Circovirus (PCV) is a known contaminant of vaccines and cell cultures that have been exposed to porcine-derived reagents. Endemic infection of PCV3 in laboratory mice has been reported, and some evidence supports natural PCV infection in wild mice. PPE samples from 2 different vendors tested positive for DNA from both PCV2 and 3. To allow model development with these reagents to proceed, we developed a protocol that would meet scientific objectives, minimize exposure of mice, and provide information on the potential for the virus to spread. Five d after BALB/c mice received intralaryngeal administration of PPE, lungs were harvested and analyzed for evidence of disease. Tissues from other major organs were submitted to test for disseminated PCV2 and 3 DNA. Similarly, tissues (including lungs) from direct contact nude sentinel mice were analyzed for the presence of the virus. To evaluate the possibility of endemic PCV2/3 infection, we also surveyed non-porcine reagent exposed mice on other studies. PCV2 and 3 was not detected in any of the tissues submitted. Although this study provided no evidence of infection and transmission of PCV2/3 from the contaminated PPE sample over the 5 d study, further work is needed to understand the risks and impact of introducing PCV contaminated cells or reagents into barrier maintained rodent colonies.

scholarly journals The Biochemical and Pathological Studies on Effects of Porcine Pancreatic Elastase in Experimental Atherosclerosis with Hyper-Cholesterolemia

1983 ◽  
Vol 11 (2) ◽  
pp. 365-374
Author(s):  
Shinzabro OHTAKE ◽  
Atsushi KOIDE ◽  
Hiroyuki SHIOJIRI ◽  
Kouichi KATAYAMA ◽  
Seiichi KOBAYASHI ◽  
...  
Keyword(s):  
Experimental Atherosclerosis ◽  
Pancreatic Elastase ◽  
Porcine Pancreatic Elastase

Rapid screening for deficiency of alpha 1-proteinase inhibitor.

1989 ◽  
Vol 35 (9) ◽  
pp. 1971-1975 ◽  
Author(s):  
C Lloyd ◽  
J Travis
Keyword(s):  
Human Plasma ◽  
Inhibitory Activity ◽  
Proteinase Inhibitor ◽  
Rapid Screening ◽  
Early Screening ◽  
Pancreatic Elastase ◽  
Low Concentrations ◽  
Normal Individuals ◽  
Porcine Pancreatic Elastase ◽  
Alpha 1 Antitrypsin

Abstract This rapid screening procedure for detection of low but functional elastase-inhibitory activity in human plasma is based on the fact that incubation of excess porcine pancreatic elastase (EC 3.4.21.36) with plasma results in formation of a complex with active alpha 1-proteinase inhibitor (alpha 1PI, also called alpha 1-antitrypsin). In normal individuals all of the elastase is complexed, leaving no free enzyme to hydrolyze the elastase substrate, and the reaction mixture remains clear. Because individuals homozygous for the Z allele have relatively low concentrations of alpha 1PI, their plasma cannot complex all of the elastase in the assay. The uncomplexed enzyme hydrolyzes the elastase-specific p-nitroanilide substrate, producing a yellow reaction mixture. Use of this simple assay for early screening of individuals for alpha 1PI deficiency may substantially decrease the number of untreated cases of familial emphysema, a disorder that develops as a result of a genetically derived proteinase-proteinase inhibitor imbalance.

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