C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/48 (2006.01) C12Q 1/18 (2006.01) G01N 33/573 (2006.01)
Patent
CA 2232173
The present invention relates to screening methods for the identification of compounds and compositions useful as novel antibiotics and antibacterial agents. In particular, the present invention relates to methods utilizing two- component regulatory switches, for example, those comprising a prokaryotic enzyme such as histidine protein kinase that is activated to autophosphorylate by a signal transduction mechanism. The invention also relates to methods of identifying inhibitors of enzyme activity, particularly in bacterial cells. In particular, a high-throughput assay system useful in the large-scale screening of protein kinase inhibitors is disclosed. Related kits, compositions and components are also disclosed. The figure illustrates the phosphorylation of SpoOF including a histidine linker or tag ("His-SpoOF") by KinA.
La présente invention se rapporte à des procédés de criblage s'appliquant à l'identification de nouveaux antibiotiques et à des compositions utilisées comme nouveaux antibiotiques et agents antibactériens. La présente invention se rapporte notamment à des procédés mettant en oeuvre des éléments de déclenchement régulateurs à deux composés, par exemple, ceux comprenant une enzyme procaryotique telle que la protéine kinase de l'histidine qui subit une autophosphorylation par un mécanisme de transduction de signaux. L'invention se rapporte également à des procédés d'identification d'inhibiteurs de l'activité enzymatique, notamment dans des cellules bactériennes, et en outre, à un système de dosage du débit utilisé dans le criblage à grande échelle des inhibiteurs de la protéine kinase. Des kits, compositions et composants connexes sont également décrits. La figure illustre la phosphorylation par KinA de SpoOF comprenant un élément de liaison histidine ou étiquette ("His-SpoOF").
Hoch James A.
Huang Shaoming
Fetherstonhaugh & Co.
The Scripps Research Institute
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