Screening methods for the identification of novel antibiotics
AssigneeThe Scripps Research Institute
InventorsJames A. Hoch, Shaoming Huang
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Abstract
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.
Claims16
An assay method for identifying antibiotic or antimicrobial agents, comprising:
- a. affixing a substrate onto a solid support;
- b. admixing said solid phase-affixed substrate with a sensor protein, a high-energy phosphate source, and a test sample, wherein said substrate comprises a bacterial protein or polypeptide molecule that includes one or more histidine residues attached to its N-terminus;
- c. allowing said admixture to incubate for a predetermined period of time;
- and
- d. examining said admixture after said incubation in order to determine the effect of said test sample upon the activity of said sensor protein.
A high-throughput assay system for screening protein kinase inhibitors comprising, in separate containers:
- a. a substrate affixed to a solid support, wherein said substrate comprises a bacterial protein or a polypeptide molecule that includes one or more histidine residues attached to its N-terminus;
- b. a sensor protein;
- and
- c. a high-energy phosphate source, each in an amount sufficient to conduct at least one assay.
Dependencies are inferred from the claim text.