Genetic and structural validation of Aspergillus fumigatus UDP-N-acetylglucosamine pyrophosphorylase as a potential antifungal target

Fang, Wenxia and Du, Ting and Raimi, Olawale G. and Hurtado-Guerrero, Ramon and Urbaniak, Mick and Ibrahim, Adel F. M. and Ferguson, Michael A. J. and van Aalten, Daan M. F. (2013) Genetic and structural validation of Aspergillus fumigatus UDP-N-acetylglucosamine pyrophosphorylase as a potential antifungal target. Molecular Microbiology, 89 (3). pp. 479-493. ISSN 0950-382X

[img]
Preview
PDF (Fang-MolMicro-2013)
Fang_MolMicro_2013.pdf - Published Version
Available under License Creative Commons Attribution.

Download (1MB)

Abstract

The sugar nucleotide UDP-N-acetylglucosamine (UDP-GlcNAc) is an essential metabolite in both prokaryotes and eukaryotes. In fungi, it is the precursor for the synthesis of chitin, an essential component of the fungal cell wall. UDP-N-acetylglucosamine pyrophosphorylase (UAP) is the final enzyme in eukaryotic UDP-GlcNAc biosynthesis, converting UTP andN-acetylglucosamine-1-phosphate (GlcNAc-1P) to UDP-GlcNAc. As such, this enzyme may provide an attractive target against pathogenic fungi. Here, we demonstrate that the fungal pathogen Aspergillus fumigatus possesses an active UAP (AfUAP1) that shows selectivity for GlcNAc-1P as the phosphosugar substrate. A conditional mutant, constructed by replacing the native promoter of the A. fumigatus uap1 gene with the Aspergillus nidulans alcA promoter, revealed that uap1 is essential for cell survival and important for cell wall synthesis and morphogenesis. The crystal structure of AfUAP1 was determined and revealed exploitable differences in the active site compared with the human enzyme. Thus AfUAP1 could represent a novel antifungal target and this work will assist the future discovery of small molecule inhibitors against this enzyme.

Item Type:
Journal Article
Journal or Publication Title:
Molecular Microbiology
Additional Information:
© 2013 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/2400/2404
Subjects:
ID Code:
66386
Deposited By:
Deposited On:
17 Sep 2013 08:07
Refereed?:
Yes
Published?:
Published
Last Modified:
03 Jun 2020 01:59