Assay for inhibitors of ispH

Details Assay for inhibitors of ispH Assay for inhibitors of ispH

2004-01-15

2008-07-22

Weber, Jon P (Department: 1657)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving oxidoreductase

Reexamination Certificate

active

07402408

ABSTRACT:
Assay for testing a sample for the presence or absence of inhibition of the enzymatic conversion of 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate into isopentenyl diphosphate and/or dimethylallyl diphosphate by the following steps: (a) reacting an aqueous mixture containing 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate, a 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphnte reductase, and a reducing agent under predetermined reaction conditions for a predetermined period of time; (b) analyzing the reaction mixture obtained in step (a) for the consumed amount of 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate and/or NAD(P)H and/or for the produced amount of isopentenyl diphosphate, and/or dimethylallyl diphosphate and/or NAD(P)+; (c) repeating step (a) in the presence of the sample to be tested; (d) repeating step (b) with the reaction mixture defined in step (c); (e) comparing the results of steps (b) and (d).

REFERENCES:
patent: WO-02/083720 (2002-10-01), None
patent: WO-2004/035810 (2004-04-01), None
Ollagnier, S. et al., “Activation Of The Anaerobic Ribonucleotide Reductase FromEscherichia coli, The Essential Role Of The Iron-Sulfur Center ForS-Adenosylmethionine Reduction”, The Journal of Biological Chemistry 272 (39) (1997), pp. 24216-24223.
Adam, P. et al., “Biosynthesis Of Terpenes: Studies On 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate Reductase”, Proceedings of The National Academy of Sciences of the United States of America, 99 (19) Sep. 17, 2002, pp. 12108-12113.
Altincicek, B. et al., “LytB Protein Catalyzes The Terminal Step Of The 2-C-methyl-D-erythritol-4-phosphate Pathway Of Isoprenoid Biosynthesis”, FEBS Letters 532 (2002), pp. 437-440.
Rohdich, F., et al. “The Deoxyxylose Phosphate Pathway of Isoprenoid Biosynthesis: Studies on the Mechanisms of the Reactions Catalyzed by IspG and IspH protein,” PNAS, vol. 100, No. 4, 2003, pp. 1586-1591.

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