A high-throughput screen utilizing the fluorescence of riboflavin for identification of lumazine synthase inhibitors

Jinhua Chen, Boris Illarionov, Adelbert Bacher, Markus Fischer, Ilka Haase, Gunda Georg, Qizhuang Ye, Zeqiang Ma, Mark Cushman

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

A high-throughput screening method based on the competitive binding of a lumazine synthase inhibitor and riboflavin to the active site of Schizosaccharomyces pombe lumazine synthase was developed. This assay is sensitive, simple, and robust. During assay development, all of the known active inhibitors tested were positively identified. Preliminary high-throughput screening in 384-well format resulted in a Z factor of 0.7. The approach utilizes a thermodynamic assay to bypass the problems associated with the instabilities of both lumazine synthase substrates that complicate the use of a kinetic assay in a high-throughput format, and it removes the time element from the assay, thus simplifying the procedure.

Original languageEnglish (US)
Pages (from-to)124-130
Number of pages7
JournalAnalytical Biochemistry
Volume338
Issue number1
DOIs
StatePublished - Mar 1 2005
Externally publishedYes

Fingerprint

Riboflavin
Assays
Fluorescence
Throughput
High-Throughput Screening Assays
Competitive Binding
Schizosaccharomyces
Screening
Thermodynamics
Catalytic Domain
6,7-dimethyl-8-ribityllumazine synthase
Kinetics
Substrates

Keywords

  • Enzyme inhibitor discovery
  • High-throughput screening
  • Lumazine synthase
  • Riboflavin synthase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A high-throughput screen utilizing the fluorescence of riboflavin for identification of lumazine synthase inhibitors. / Chen, Jinhua; Illarionov, Boris; Bacher, Adelbert; Fischer, Markus; Haase, Ilka; Georg, Gunda; Ye, Qizhuang; Ma, Zeqiang; Cushman, Mark.

In: Analytical Biochemistry, Vol. 338, No. 1, 01.03.2005, p. 124-130.

Research output: Contribution to journalArticle

Chen, J, Illarionov, B, Bacher, A, Fischer, M, Haase, I, Georg, G, Ye, Q, Ma, Z & Cushman, M 2005, 'A high-throughput screen utilizing the fluorescence of riboflavin for identification of lumazine synthase inhibitors', Analytical Biochemistry, vol. 338, no. 1, pp. 124-130. https://doi.org/10.1016/j.ab.2004.11.033
Chen, Jinhua ; Illarionov, Boris ; Bacher, Adelbert ; Fischer, Markus ; Haase, Ilka ; Georg, Gunda ; Ye, Qizhuang ; Ma, Zeqiang ; Cushman, Mark. / A high-throughput screen utilizing the fluorescence of riboflavin for identification of lumazine synthase inhibitors. In: Analytical Biochemistry. 2005 ; Vol. 338, No. 1. pp. 124-130.
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