Detection of human leukocyte elastase from a plasma alpha-1-proteinase inhibitor complex by its enzymatic activity with a synthetic substrate

A spectrophotometric procedure was developed for estimation of elastase activity in human leukocytes in the form of complex with blood serum alpha 1 proteinase inhibitor after loosening of the complex and detection of the enzymatic activity with N-tert-butyl-hydroxy-carbonyl-Ala-p-nitrophenyl ester...

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Bibliographic Details
Published inVoprosy meditsinskoi khimii Vol. 40; no. 3; p. 20
Main Authors Dotsenko, V L, Neshkova, E A, Iarovaia, G A
Format Journal Article
LanguageRussian
Published Russia (Federation) 01.05.1994
Subjects
Acetone
Acetonitriles
alpha 1-Antitrypsin - metabolism
Chromatography, Ion Exchange
Humans
Hydrogen-Ion Concentration
Leukocyte Elastase
Pancreatic Elastase - antagonists & inhibitors
Pancreatic Elastase - isolation & purification
Pancreatic Elastase - metabolism
Spectrum Analysis
Substrate Specificity
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Summary:A spectrophotometric procedure was developed for estimation of elastase activity in human leukocytes in the form of complex with blood serum alpha 1 proteinase inhibitor after loosening of the complex and detection of the enzymatic activity with N-tert-butyl-hydroxy-carbonyl-Ala-p-nitrophenyl ester as a substrate in presence of acetone or acetonitrile. The optimal conditions were described, which were required for estimation of the enzyme 100% activity followed by addition of the leukocyte elastase standard preparation into blood serum as well as for measurement of the enzyme activity in the complex with alpha 1 proteinase inhibitor. The procedure took 6-10 min to evaluate the elastase activity in the complex with the inhibitor using simple buffer containing organic solvents at 30 degrees and the available two-beam spectrophotometer.
ISSN:0042-8809