10.1021/bc200346e.s001
Ying Zhao
Ying
Zhao
Dalia Hammoudeh
Dalia
Hammoudeh
Wenwei Lin
Wenwei
Lin
Sourav Das
Sourav
Das
Mi-Kyung Yun
Mi-Kyung
Yun
Zhenmei Li
Zhenmei
Li
Elizabeth Griffith
Elizabeth
Griffith
Taosheng Chen
Taosheng
Chen
Stephen W. White
Stephen W.
White
Richard E. Lee
Richard E.
Lee
Development of a Pterin-Based Fluorescent Probe for Screening Dihydropteroate Synthase
American Chemical Society
2011
sulfonamide class
acid binding pocket
pterin
DHPS FP assay
novel DHPS inhibitors
Screening Dihydropteroate SynthaseDihydropteroate synthase
phosphate release assays
DMSO
2011-10-19 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Development_of_a_Pterin_Based_Fluorescent_Probe_for_Screening_Dihydropteroate_Synthase/2599159
Dihydropteroate synthase (DHPS) is the classical target of the sulfonamide class of antimicrobial agents, whose use has been limited by widespread resistance and pharmacological side effects. We have initiated a structure-based drug design approach for the development of novel DHPS inhibitors that bind to the highly conserved and structured pterin subsite rather than to the adjacent <i>p</i>-aminobenzoic acid binding pocket that is targeted by the sulfonamide class of antibiotics. To facilitate these studies, a robust pterin site-specific fluorescence polarization (FP) assay has been developed and is discussed herein. These studies include the design, synthesis, and characterization of two fluorescent probes, and the development and validation of a rapid DHPS FP assay. This assay has excellent DMSO tolerance and is highly reproducible as evidenced by a high Z′ factor. This assay offers significant advantages over traditional radiometric or phosphate release assays against this target, and is suitable for site-specific high-throughput and fragment-based screening studies.