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Genetic and structural validation of A spergillus fumigatus UDP ‐ N ‐acetylglucosamine pyrophosphorylase as an antifungal target
Author(s) -
Fang Wenxia,
Du Ting,
Raimi Olawale G.,
HurtadoGuerrero Ramon,
Urbaniak Michael D.,
Ibrahim Adel F. M.,
Ferguson Michael A. J.,
Jin Cheng,
Aalten Daan M. F.
Publication year - 2013
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.12290
Subject(s) - aspergillus fumigatus , biology , n acetylglucosamine , enzyme , chitin , biochemistry , aspergillus nidulans , cell wall , mutant , conidiation , pathogen , biosynthesis , microbiology and biotechnology , gene , chitosan
Summary 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. U DP ‐ N ‐ a cetylglucosamine p yrophosphorylase ( UAP ) is the final enzyme in eukaryotic UDP ‐ GlcNAc biosynthesis, converting UTP and N ‐acetylglucosamine‐1‐phosphate ( GlcNAc ‐1 P ) to UDP ‐ GlcNAc . As such, this enzyme may provide an attractive target against pathogenic fungi. Here, we demonstrate that the fungal pathogen A spergillus fumigatus possesses an active UAP ( Af UAP 1) that shows selectivity for GlcNAc ‐1 P as the phosphosugar substrate. A conditional mutant, constructed by replacing the native promoter of the A . fumigatus uap1 gene with the A spergillus nidulans alcA promoter, revealed that uap1 is essential for cell survival and important for cell wall synthesis and morphogenesis. The crystal structure of Af UAP 1 was determined and revealed exploitable differences in the active site compared with the human enzyme. Thus Af UAP 1 could represent a novel antifungal target and this work will assist the future discovery of small molecule inhibitors against this enzyme.