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US 2003/0143635 A1Published application
Assay for detection of transferase enzyme activity in drug screening
AssigneeAstraZeneca AB
InventorsSunita Maria Desousa, Suresh Solapure
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Abstract
The invention provides methods for assaying the activity of the translocase enzyme and/or transferase enzyme involved in peptidoglycan biosynthesis in bacteria using scintillation proximity assay methodology. The methods are suitable for high throughput screening of potential anti-bacterial drugs.
Claims12
A method for assaying UDP-N-acetylglucosamine:
- N-acetylmuramyl(pentapeptide)-P-P-undecaprenol-N-acetylglucosamine transferase enzyme activity, and, optionally also phospho-N-acetylmuramyl-pentapeptide translocase enzyme activity, which method comprises the steps of:
- (1) incubating a reaction mixture comprising undecaprenol-pyrophosphate-N-acetylmuramylpentapeptide (Lipid I), radiolabelled UDP-N-acetyl glucosamine (UDP-GlcNAc), a source of divalent metal ions and a source of the transferase enzyme under conditions suitable for synthesis of undecaprenol-pyrophosphoryl-N-acetylmuramylpentapeptide-N-acetylglucosamine (Lipid II) to occur;
- (2) stopping the reaction of step (1);
- (3) adding to the reaction mixture of step (2) a fluorescer;
- and
- (4) measuring light energy emitted by the fluorescer.
3 more elements
Dependencies are inferred from the claim text.