Probing cellular protein targets of H2O2 with fluorescein-conjugated iodoacetamide and antibodies to fluorescein

被引：83

作者：

Wu, YL ^{[1
]}

Kwon, KS ^{[1
]}

Rhee, SG ^{[1
]}

机构：

[1] NHLBI, Lab Cell Signaling, NIH, Bethesda, MD 20892 USA

来源：

FEBS LETTERS | 1998年 / 440卷 / 1-2期

关键词：

hydrogen peroxide; intracellular messenger; cysteine oxidation; growth factor; protein tyrosine phosphatase;

D O I：

10.1016/S0014-5793(98)01415-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Recent studies suggest that H2O2, at subtoxic concentrations generated in response to the activation of a variety of cell surface receptors, functions as an intracellular messenger. However the intracellular targets of H2O2 action have not; been identified. A procedure to detect proteins with reactive cysteine residues susceptible to oxidation by intracellularly generated H2O2 is now described, This approach is based on the labeling of proteinaceous cysteine with 5-iodoacetamido-fluorescein at pH 5.5 and immunoblot analysis of the labeled proteins with antibodies specific to fluorescein. With this procedure, many proteins in human A431 cells were shown to contain reactive cysteines and to be readily oxidized by H2O2 generated in response to cellular stimulation with epidermal growth factor. One of these H2O2-sensitive proteins was identified as protein tyrosine phosphatase 1B. (C) 1998 Federation of European Biochemical Societies.

引用

页码：111 / 115

页数：5

共 19 条

[1] Epidermal growth factor (EGF)-induced generation of hydrogen peroxide - Role in EGF receptor-mediated tyrosine phosphorylation [J].