Covalent tethering of functional groups to proteins and...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/62 (2006.01) A01K 67/027 (2006.01) A61K 38/00 (2006.01) A61K 38/46 (2006.01) A61K 49/00 (2006.01) B01D 57/00 (2006.01) C07D 311/80 (2006.01) C07D 407/12 (2006.01) C07D 495/04 (2006.01) C07H 21/00 (2006.01) C07K 1/13 (2006.01) C07K 1/14 (2006.01) C07K 1/22 (2006.01) C07K 14/195 (2006.01) C12N 1/00 (2006.01) C12N 5/00 (2006.01) C12N 9/14 (2006.01) C12N 11/00 (2006.01) C12N 15/55 (2006.01) C12N 15/85 (2006.01) C12Q 1/00 (2006.01) C12Q 1/34 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01) G01N 33/566

Patent

CA 2575611

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

L'invention concerne une hydrolase mutante facultativement fusionnée à une protéine d'intérêt. Ladite hydrolase mutante est capable de former une liaison avec un substrat en vue de l'hydrolase non mutante correspondante (de type sauvage) plus stable que la liaison formée entre l'hydrolase de type sauvage et le substrat et présente au moins deux substitutions d'acides aminés par rapport à l'hydrolase de type sauvage. Cette invention a aussi pour objet des substrats destinés à des hydrolases comprenant au moins un groupe fonctionnel, des méthodes d'utilisation de l'hydrolase mutante et des substrats de ladite invention, ainsi qu'une protéine hybride capable de former une liaison stable avec un substrat et des cellules qui expriment la protéine hybride.

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