CONTINUOUS AND DISCONTINUOUS ASSAYS FOR PHOSPHOTYROSYL PROTEIN PHOSPHATASE-ACTIVITY USING PHOSPHOTYROSYL PEPTIDE-SUBSTRATES

被引：28

作者：

NASH, K

FELDMULLER, M

DEJERSEY, J

ALEWOOD, P

HAMILTON, S

机构：

[1] UNIV QUEENSLAND, DEPT BIOCHEM, ST LUCIA, BRISBANE 4072, AUSTRALIA

[2] UNIV QUEENSLAND, CTR DRUG DESIGN & DEV, ST LUCIA, BRISBANE 4072, AUSTRALIA

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 213卷 / 02期

关键词：

D O I：

10.1006/abio.1993.1425

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Assays for phosphotyrosyl protein phosphatase activity have been developed using the synthetic peptide Glu-Glu-Tyr(P)-Ala-Ala and its N(α)-fluorenylmethoxycarbonyl derivative (Fmoc-Glu-Glu-Tyr(P)-Ala-Ala) as substrates. Conditions for monitoring the phosphatase-catalyzed hydrolysis of the former peptide, either spectrophotometrically or fluorometrically, are reported. These continuous assays are similar in sensitivity to published assays using phosphotyrosine as substrate. A discontinuous HPLC-based assay using the intensely fluorescent Fmoc-peptide as substrate is also described. This assay is comparable in sensitivity to assays using 32P-labeled substrates and is suitable for assaying low levels of phosphotyrosyl protein phosphatase activity in crude tissue extracts. © 1994 Academic Press, Inc. All rights reserved.

引用

页码：303 / 309

页数：7

共 22 条

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