[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 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.
Dotsenko
V L
VL
Neshkova
E A
EA
Iarovaia
G A
GA
D000096:Acetone; D000097:Acetonitriles; D002852:Chromatography, Ion Exchange; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019272:Leukocyte Elastase; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D013057:Spectrum Analysis; D013379:Substrate Specificity; D000515:alpha 1-Antitrypsin / Q000378:metabolism*
D000097:Acetonitriles; D000515:alpha 1-Antitrypsin; D000096:Acetone; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase; C032159:acetonitrile
Vyiavlenie leĭkotsitarnoĭ élastazy cheloveka iz kompleksa s plazmennym alph1-proteinaznym ingibitorom po ee énzimaticheskoĭ aktivnosti s sinteticheskim substratom.