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Biological Procedures Online

, Volume 4, Issue 1, pp 1–9

Received: 26 March 2002Accepted: 16 May 2002DOI: 10.1251-bpo28

Cite this article as: Zhu, L. & Goldstein, B. Biol Proced Online 2002 4: 1. doi:10.1251-bpo28

Abstract

Protein-tyrosine phosphatases PTPases have a catalytic cysteine residue whose reduced state is integral to the reaction mechanism. Since exposure to air can artifactually oxidize this highly reactive thiol, PTPase assays have typically used potent reducing agents to reactivate the enzymes present; however, this approach does not allow for the measurement of the endogenous PTPase activity directly isolated from the in vivo cellular environment. Here we provide a method for using an anaerobic chamber to preserve the activity of the total PTPase complement in a tissue lysate or of an immunoprecipitated PTPase homolog to characterize their endogenous activation state. Comparison with a sample treated with biochemical reducing agents allows the determination of the activatable reducible fraction of the endogenous PTPase pool.

Indexing termssignal transduction cysteine protein-tyrosine-phosphatase oxidation-reduction Published: June 11, 2002.

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Autor: Li Zhu - Barry Goldstein

Fuente: https://link.springer.com/







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