Engineered protein kinases which can utilize modified...

C - Chemistry – Metallurgy – 07 – H

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C07H 19/167 (2006.01) A61K 38/45 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12N 15/09 (2006.01) C12N 15/54 (2006.01) C12Q 1/00 (2006.01) C12Q 1/02 (2006.01) C12Q 1/48 (2006.01) G01N 33/50 (2006.01) G01N 33/573 (2006.01) G01N 33/60 (2006.01)

Patent

CA 2279846

Engineered protein kinases which can utilize modified nucleotide triphosphate substrates that are not as readily utilized by the wild-type forms of those enzymes, and methods of making and using them. Modified nucleotide triphosphate substrates and methods of making and using them. Methods for using such engineered kinases and such modified substrates to identify which protein substrates the kinases act upon, to measure the extent of such action, and to determine if test compounds can modulate such action. Also engineered forms of multi-substrate enzymes which covalently attach part or all of at least one (donor) substrate to at least one other (recipient) substrate, which engineered forms will accept modified substrates that are not as readily utilized by the wild-type forms of those enzymes. Methods for making and using such engineered enzymes. Modified substrates and methods of making and using them. Methods for using such engineered enzymes and such modified substrates to identify the recipient substrates the enzymes act upon, to measure the extent of such action, and to measure whether test compounds modulate such action.

L'invention concerne des protéines kinases manipulées pouvant utiliser des substrats triphosphate nucléotidique modifiés qui ne sont pas aussi facilement utilisés par les formes de type sauvage de ces enzymes, et leurs procédés de production et d'utilisation; des substrats triphosphate nucléotidique modifiés et leurs procédés de production et d'utilisation; des procédés d'utilisation de telles kinases manipulées et de tels substrats modifiés, pour identifier les substrats protéiques sur lesquels agissent les kinases, pour mesurer l'étendue d'une telle action et pour déterminer si les composés d'essai sont susceptibles de moduler cette action. L'invention concerne également des formes manipulées d'enzymes multi-substrats liant, de façon covalente, une partie ou la totalité d'au moins un substrat (donneur) à un autre substrat (receveur) au moins, dont les formes manipulées acceptent des substrats modifiés qui ne sont pas aussi facilement utilisés par les formes de type sauvage de ces enzymes; des procédés de production et d'utilisations de tels enzymes manipulées; des substrats modifiés et leurs procédés de production et d'utilisation; des procédés d'utilisation de tels enzymes manipulées et de tels substrats modifiés pour identifier les substrats receveurs sur lesquels agissent les enzymes, pour mesurer l'étendue d'une telle action et pour déterminer si les composés d'essai modulent cette action.

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