Methods for screening inhibitors of apoptosis

Details Methods for screening inhibitors of apoptosis Methods for screening inhibitors of apoptosis

C - Chemistry, Metallurgy

12

Q

C12Q 1/37 (2006.01)

Patent

CA 2526515

The present invention addresses a need in the art for methods of identifying apoptotic proteins and methods for screnning compounds which inhibit an poptotic protein. More particularly, in certain embodiments, the invention relates to increased expression levels of a NALP1 gene and/or a NALP5 gene following neuron injury. In other embodiments, the present invention demonstrates that the recombinant expression of NALP1 and/or NALP polynucleotides stimulates apoptosis in cultured neurons, Hela cells and NIH- 3T3 cells. In yet other embodiments, the invention relates to mutations in the nucleotide binding sequence (NBS) of a NALP or a NALP5 polypeptide, wherein these NBS mutations inhibit purine nucleotide binding and reduce caspase activation.

L'invention concerne le besoin dans la technique antérieure de méthodes d'identification de protéines apoptotiques et de méthodes de criblage de composés inhibant une protéine apoptotique. Dans certains modes de réalisation, l'invention porte notamment sur des niveaux d'expression accrus d'un gène NALP1 et/ou d'un gène NALP5 après une lésion neuronale. Dans d'autres modes de réalisation, il est démontré que l'expression de recombinaison des polynucléotides NALP1 et/ou NALP5 stimule l'apoptose dans des neurones mis en culture, à savoir les cellules HeLa et les cellules NIH-3T3. Dans encore d'autres modes de réalisation, l'invention se rapporte à des mutations de la séquence de liaison nucléotidique (NBS) d'un polypeptide NALP1 ou NALP5, ces mutations de NBS inhibant la liaison nucléotidique purine et réduisent l'activation de la caspase.

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