Protein engineering of monoacylglycerol lipase (mgll)

Details Protein engineering of monoacylglycerol lipase (mgll) Protein engineering of monoacylglycerol lipase (mgll)

C - Chemistry, Metallurgy

12

N

C12N 9/18 (2006.01) C12Q 1/44 (2006.01)

Patent

CA 2722446

A number of soluble engineered forms of MGLL that are suitable for high- throughput screening and protein crys-tallization, as well as a crystallized form of monoacylglycerol lipase protein (MGLL) and descriptions of the X-ray diffraction pat-terns are disclosed. The engineered constructs of MGLL permit the expression and purification of protein suitable for crystallogra-phy or high-throughput screening and identification of ligands, which can function as active agents to MGLL. The X-ray diffrac-tion patterns allow the three dimensional structure of MGLL to be determined at atomic resolution so that ligand binding sites on MGLL can be identified and the interactions of ligands with MGLL amino acid residues can be modeled. Models prepared using such maps permit the design of ligands which can function as active agents which include, but are not limited to, those that func-tion as inhibitors of MGLL.

Linvention concerne un certain nombre de formes conçues solubles de MGLL qui sont appropriées pour un criblage à haut débit et la cristallisation de protéine, ainsi quune forme cristallisée de protéine de monoacylglycérol lipase (MGLL) et des descriptions des diagrammes de diffraction aux rayons X. Les produits de construction conçus de MGLL permettent lexpression et la purification de protéines appropriées pour une cristallographie ou un criblage à haut débit et lidentification de ligands, qui peuvent fonctionner comme des agents actifs à la MGLL. Les diagrammes de diffraction aux rayons X permettent à la structure tridimensionnelle de MGLL dêtre déterminée à une résolution atomique de sorte que des sites de liaison de ligand sur la MGLL peuvent être identifiés et les interactions de ligands avec les résidus dacides aminés de MGLL peuvent être modélisées. Des modèles préparés à laide de telles cartes permettent à la conception de ligands qui peuvent fonctionner comme des agents actifs qui comprennent, sans s'y limiter, ceux qui fonctionnent comme des inhibiteurs de MGLL.

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