WATER-SOLUBLE MACROMOLECULAR FLUOROGENIC SUBSTRATES FOR ASSAYING PROTEINASES - DETERMINATION OF PANCREATIC ELASTASE ACTIVITY

被引：6
|
作者：
ANJUERE, F
MONSIGNY, M
MAYER, R
机构：
[1] Département de Biochimie des Glycoconjugués et des Lectines Endogènes, Centre de Biophysique Moléculaire, CNRS, 45071 Orléans Cedex 02
来源：
ANALYTICAL BIOCHEMISTRY | 1991年 / 198卷 / 02期
关键词：
D O I：
10.1016/0003-2697(91)90436-W
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Water-soluble macromolecular fluorogenic substrates were synthesized in order to develop an easy specific proteinase assay. The validity of this method was tested with porcine pancreatic elastase by using its specific peptidic substrate Ala-Ala-Pro-Ala linked to a hydrosoluble polymer. The octapeptidic sequence FTC-ε{lunate}Aca-Ala-Ala-Pro-Ala-Gly-Gly-Gly was linked to a water-soluble and neutral poly-l-lysine derivative. The aminocaproyl residue and the triglycyl sequence were added in order to improve the stability of the substrate, and the accessibility of the specific sequence Ala-Ala-Pro-Ala to elastase, respectively. The assay is based on the quantitative precipitation of the polymeric substrate in isopropanol while the released soluble fluorescent peptidic moiety is fluorometrically titrated in the supernatant. © 1991.
引用
收藏
页码：342 / 346
页数：5
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