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[通过其与合成底物的酶活性检测血浆α-1-蛋白酶抑制剂复合物中的人白细胞弹性蛋白酶]

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

作者信息

Dotsenko V L, Neshkova E A, Iarovaia G A

出版信息

Vopr Med Khim. 1994 May-Jun;40(3):20-5.

PMID:8079434
Abstract

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.

摘要

开发了一种分光光度法,用于在复合物解离后,以与血清α1蛋白酶抑制剂形成的复合物形式估计人白细胞中的弹性蛋白酶活性,并在丙酮或乙腈存在下,以N-叔丁基-羟基-羰基-Ala-对硝基苯酯为底物检测酶活性。描述了最佳条件,在向血清中添加白细胞弹性蛋白酶标准制剂以估计酶的100%活性以及测量与α1蛋白酶抑制剂形成的复合物中的酶活性时都需要这些条件。使用含有有机溶剂的简单缓冲液,在30摄氏度下,利用现有的双光束分光光度计,该方法需要6-10分钟来评估与抑制剂形成的复合物中的弹性蛋白酶活性。

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