Evaluation of recombinant caspase specificity by competitive substrates

被引：34

作者：

Benkova, Bistra ^{[1
]}

Lozanov, Valentin ^{[1
]}

Ivanov, Ivaylo P. ^{[2
]}

Mitev, Vanio ^{[1
]}

机构：

[1] Med Univ Sofia, Dept Med Chem & Biochem, Sofia 1431, Bulgaria

[2] Univ Sofia, Fac Biol, Sofia 1164, Bulgaria

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 394卷 / 01期

关键词：

Competitive substrates; Fluorescent substrates; Caspase specificity; 2-Aminoacridone; HPLC; COMBINATORIAL APPROACH; CONVERTING-ENZYME; ACTIVATION; FAMILY;

D O I：

10.1016/j.ab.2009.07.012

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

The specificity of 10 recombinant caspases was investigated using a set of competitive substrates. The caspase activity was determined by high-performance liquid chromatography using highly fluorescent peptides containing 2-aminoacridone (AMAC) as reporting group. The sequences of the used substrates were designed according to literature data for being specific for 10 of the caspases. The described approach allows the concentration changes of several substrates to be monitored simultaneously in a single sample. Because the substrates are in competitive conditions, the preferences of particular caspases to given peptide sequences are most clearly demonstrated. In the studied competitive assay conditions, all tested caspases except caspase 2 exhibit activity toward more than one substrate. None of the used peptide sequences was found to be highly specific for a defined caspase. The results obtained indicate that there is well-expressed group specificity among the caspases. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：68 / 74

页数：7

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