A versatile spectrophotometric protein tyrosine phosphatase assay based on 3-nitrophosphotyrosine containing substrates

Jeroen van Ameijde, John Overvoorde, Stefan Knapp, Jeroen den Hertog, Rob Ruijtenbeek, Rob M J Liskamp

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

Samenvatting

A versatile assay for protein tyrosine phosphatases (PTP) employing 3-nitrophosphotyrosine containing peptidic substrates is described. These therapeutically important phosphatases feature in signal transduction pathways. The assay involves spectrophotometric detection of 3-nitrotyrosine production from 3-nitrophosphotyrosine containing peptidic substrates, which are accepted by many PTPs. Compared to conventional chromogenic phosphate derivatives, the more realistic peptidic substrates allow evaluating substrate specificity. The assay's applicability is demonstrated by determining kinetic parameters for several PTP-substrate combinations and inhibitor evaluation, as well as detection of PTP activity in lysates. The convenient new assay may assist further adoption of PTPs in drug development.

Originele taal-2Engels
Pagina's (van-tot)9-13
Aantal pagina's5
TijdschriftAnalytical Biochemistry
Volume448
Vroegere onlinedatum03 dec 2013
DOI's
StatusGepubliceerd - 01 mrt 2014

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