Cytotoxic cyplasin of the sea hare, aplysia punctata, cdna...

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/12 (2006.01) A61K 31/7088 (2006.01) A61K 38/17 (2006.01) A61K 48/00 (2006.01) A61P 35/00 (2006.01) C07K 14/435 (2006.01) C12N 5/10 (2006.01) C12N 15/79 (2006.01) C12P 21/02 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2472280

Described is a nucleic acid coding for a protein called "cyplasin" that shows a preferential toxicity to autonomously growing mammalian cells. Cell death induced by this protein differs from both apoptosis and necrosis. An intracellular cell death which occurs when recombinantly preparing cyplasin in cell cultures can be avoided by removal of the secretion signal in the cyplasin sequence. This modification makes it possible to express the cyplasin in a mammalian cell culture which is preferable with regard to the glycosylation pattern of the obtained protein. Thus, the present invention also relates to a method of recombinantly producing a protein in eukaryotic cells, preferably mammalian cells, which is cytotoxic for said cells when applied externally.

L'invention concerne un acide nucléique codant une protéine dite <= cyplasine >= qui présente une toxicité préférentielle pour les cellules de mammifères se développant de manière autonome. La mort cellulaire induite par cette protéine diffère de l'apoptose et de la nécrose. Il est possible d'éviter la mort cellulaire qui se produit au niveau intracellulaire lors de la préparation par recombinaison de cyplasine dans des cultures cellulaires en retirant le signal de secrétion dans la séquence de cyplasine. Cette modification permet d'exprimer la cyplasine dans une culture de cellules de mammifères, ce qui est préférable par rapport au schéma de glycolysation de la protéine obtenue. Ainsi, la présente invention traite également d'un procédé permettant de produire par recombinaison une protéine dans les cellules eucaryotes, de préférence des cellules de mammifères, qui est cytotoxique pour lesdites cellules lors de son application externe.

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