A high-throughput screening platform for inhibitors of the riboflavin biosynthesis pathway.
3,4-Dihydroxy-2-butanone 4-phosphate synthase, 6,7-dimethyl-8-ribityllumazine synthase, and riboflavin synthase of the riboflavin biosynthetic pathway are potential targets for novel antiinfective drugs. This article describes a platform for high-throughput screening for inhibitors of these enzymes. The assays can be monitored photometrically and have been shown to be robust, as indicated by Z factors 0.87. A (13)C NMR assay for hit verification of 3,4-dihydroxy-2-butanone 4-phosphate synthase inhibitors is also reported.
Kaiser
Johannes
J
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, D-85748 München, Germany.
Illarionov
Boris
B
Rohdich
Felix
F
Eisenreich
Wolfgang
W
Saller
Sabine
S
den Brulle
Jan Van
JV
Cushman
Mark
M
Bacher
Adelbert
A
Fischer
Markus
M
D000890:Anti-Infective Agents / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D053898:Biosynthetic Pathways; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors; D010453:Peptide Synthases / Q000037:antagonists & inhibitors; D011621:Pteridines; D012256:Riboflavin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis* / Q000737:chemistry; D012258:Riboflavin Synthase / Q000737:chemistry / Q000378:metabolism*
D000890:Anti-Infective Agents; D009097:Multienzyme Complexes; D011621:Pteridines; C001061:6,7-dimethyl-8-ribityllumazine; C044822:6,7-dimethyl-8-ribityllumazine synthase; D012258:Riboflavin Synthase; C490100:3,4-dihydroxy-2-butanone 4-phosphate synthase, C albicans; D010453:Peptide Synthases; D012256:Riboflavin