Genetic and structural validation ofA spergillus fumigatus UDP ‐N ‐acetylglucosamine pyrophosphorylase as an antifungal target
| Autor: | Ramon Hurtado-Guerrero, Cheng Jin, Olawale G. Raimi, Wenxia Fang, Michael A. J. Ferguson, Michael D. Urbaniak, Daan M. F. van Aalten, Ting Du, Adel F. M. Ibrahim |
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| Přispěvatelé: | National Natural Science Foundation of China, Wellcome Trust |
| Rok vydání: | 2013 |
| Předmět: |
Mutant
Microbiology Acetylglucosamine Substrate Specificity Aspergillus fumigatus Fungal Proteins 03 medical and health sciences chemistry.chemical_compound Chitin Cell Wall Cloning Molecular Promoter Regions Genetic Molecular Biology Gene Research Articles 030304 developmental biology chemistry.chemical_classification 0303 health sciences Aspergillus Fungal protein Genes Essential biology 030306 microbiology Active site biology.organism_classification Nucleotidyltransferases Protein Structure Tertiary 3. Good health carbohydrates (lipids) Enzyme chemistry Biochemistry Mutation biology.protein |
| Zdroj: | Molecular Microbiology Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1365-2958 0950-382X 3103-0025 |
| Popis: | 15 pags, 5 figs, 3 tabs 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 and N-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.fumigatusuap1 gene with the Aspergillus nidulansalcA 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. © 2013 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd. This work was funded by MRC Programme Grant G0900138 to D.M.F.v.A. and the National Natural Science Foundation of China (31030025) to C. Jin. D.M.F.v.A. is funded by a Wellcome Trust Senior Research Fellowship (WT087590MA) |
| Databáze: | OpenAIRE |
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