Identification of Eukaryotic UDP-galactopyranose Mutase Inhibitors using the ThermoFAD Assay

Abstract

Aspergillus fumigatus is a human pathogen responsible for deadly infections in immune-compromised patients. A potential strategy for treating A. fumigatus infections is by targeting the biosynthesis of cell wall components, such as galactofuranase, which is absent in humans. Galactofuranose biosynthesis is initiated by the flavoenzyme UDP-galactopyranose mutase (UGM), which converts UDP-galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf). UGM requires the reduced form of the flavin for activity, which is obtained by reacting with NADPH. We aimed to identify inhibitors of UGM by screening a kinase inhibitor library using ThermoFAD, a flavin fluorescence thermal shift assay. The screening assay identified flavopiridol as a compound that increased the melting temperature of A. fumigatus UGM. Further characterization showed that flavopiridol is a non-competitive inhibitor of UGM and docking studies suggest that it binds in the active site. This compound does not inhibit the prokaryotic UGM from Mycobacteria tuberculosis.

Department(s)

Chemistry

Comments

National Institute of General Medical Sciences, Grant R01GM094469

Keywords and Phrases

Aspergillus fumigatus; Flavopiridol; Galactofuran; ThermoFAD; UDP-Galactopyranose mutase; UGM inhibition

International Standard Serial Number (ISSN)

1090-2104; 0006-291X

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2024 Elsevier, All rights reserved.

Publication Date

04 Nov 2017

PubMed ID

28919416

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