A fluorescent microplate assay for microcystin-LR

被引：52

作者：

Fontal, OI

Vieytes, MR

de Sousa, JMVB

Louzao, MC

Botana, LM ^{[1
]}

机构：

[1] Univ Santiago de Compostela, Fac Vet, Dept Farmacol, Lugo 27002, Spain

[2] Univ Santiago de Compostela, Fac Vet, Dept Fisiol, Lugo 27002, Spain

[3] CECOPESCA, ANFACO, Vigo, Spain

来源：

ANALYTICAL BIOCHEMISTRY | 1999年 / 269卷 / 02期

关键词：

microcystin; okadaic acid; protein phosphatase; fluorescent assay; microplate;

D O I：

10.1006/abio.1999.3099

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A fluorescent enzyme inhibition assay for microcystin-LR was developed using a new fluorescent substrate of protein phosphatases 1 (PP1) and 2A (PP2A), 6,8-difluoro-4-methylumbelliferyl phosphate. The PP1 and PP2A inhibition assay for microcystin-LR was performed in a microtiter plate and the fluorescence yielded by the enzymatic hydrolysis of the substrate was quantified in a fluorescence plate reader. The concentration of microcystin-LR causing 50% inhibition of PP1 and PP2A activity (IC50) was 0.01 nM for PP1 and 0.08 nM for PP2A The measurable range of microcystin-LR was 800 to 0.08 pg/well for both enzymes. The described assay is fast and very sensitive for the detection of microcystin-LR. Furthermore, this assay can be successfully applied to the study of toxins that inhibit PP1 or PP2A. (C) 1999 Academic Press.

引用

页码：289 / 296

页数：8

共 33 条

[1] USE OF A COLORIMETRIC PROTEIN PHOSPHATASE INHIBITION ASSAY AND ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR THE STUDY OF MICROCYSTINS AND NODULARINS [J].