Chimeric proteins comprising a haem domain of a mammalian or...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/00 (2006.01) C12N 9/02 (2006.01) C12N 15/10 (2006.01) C12Q 1/00 (2006.01) C12Q 1/26 (2006.01)

Patent

CA 2456719

A chimeric protein comprises a haem domain from a mammalian or plant cytochrome P450 and a scaffold domain generally comprising an electron transfer domain, from P450 BM3 of Bacillus megaterium. The protein does not include the membrane binding portion of the wild type mammalian or plant P450 and is hence water soluble. The protein is enzymically active on substrates for the wild-type mammalian or plant P450 and electrons are transferred to electron transfer portions for instance part of the scaffold domain, such as FAD or FMN. The protein is useful for analysing the substrate specificity of the haem domain or for detecting substrates which are analytes of interest.

L'invention concerne une protéine chimère qui comprend un domaine hème dérivé d'un cytochrome P450 mammalien ou végétal et un domaine échafaudage comprenant généralement un domaine de transfert des électrons et dérivé de P450 BM3 du Bacillus megaterium. Cette protéine, qui ne comprend pas la partie de fixation sur la membrane du P450 mammalien ou végétal de type sauvage, est donc hydrosoluble. Cette protéine est active du point de vue enzymatique sur des substrats pour le P450 mammalien ou végétal de type sauvage et les électrons sont transférés sur des parties de transfert des électrons qui font, par exemple, partie du domaine échafaudage, telles que FAD ou FMN. Cette protéine convient pour l'analyse de la spécificité de substrat du domaine hème ou pour la détection de substrats qui sont des analytes d'intérêt.

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