Fluorescence polarization assays for...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/48 (2006.01) C12Q 1/34 (2006.01) C12N 9/10 (2006.01) C12N 9/78 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2600154

Provided are methods for determining the activity of proteins that modulate the acetylation state of a protein substrate. The methods may be used for determining both acetyltransferase activity and deacetylase activity. The methods involve fluorescence polarization measurements for determining the acetylation state of a substrate peptide. The methods may also be used to identify compounds that modulate the activity of a protein having acetyltransferase or deacetylase activity. Also provided are substrates for acetyltransferase or deacetylase enzymes for use in association with a fluorescence polarization assay.

L'invention se rapporte à des procédés de détermination de l'activité de protéines qui modulent l'état d'acétylation d'un substrat protéique. On peut utiliser ces procédés pour déterminer à la fois l'activité acétyltransférase et l'activité déacétylase. Ces procédés consistent à mesurer la polarisation de fluorescence pour déterminer l'état d'acétylation d'un peptide substrat. On peut également utiliser ces procédés pour identifier des composés qui modulent l'activité d'une protéine ayant une activité acétyltransférase ou une activité déacétylase. L'invention concerne également des substrats pour enzymes acétyltransférase ou déacétylase destinés à être utilisés en association avec un dosage par polarisation de fluorescence.

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