Characterisation of the Candida albicans Phosphopantetheinyl Transferase Ppt2 as a Potential Antifungal Drug Target
| Autor: | Sarah J. Kaye, Katharine S. Dobb, John Leslie Thain, Lubomira Stateva, Jason David Oliver, Mike Birch, Nicola Beckmann |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Antifungal Agents
Molecular Sequence Data Antifungal drug Gene Expression Transferases (Other Substituted Phosphate Groups) lcsh:Medicine Aspergillus fumigatus Microbiology law.invention Substrate Specificity 03 medical and health sciences Bacterial Proteins law Gene expression Candida albicans Amino Acid Sequence Promoter Regions Genetic lcsh:Science 030304 developmental biology Sequence Deletion 0303 health sciences Multidisciplinary biology 030306 microbiology lcsh:R Computational Biology biology.organism_classification Corpus albicans 3. Good health Enzyme Activation Acyl carrier protein Phenotype Biochemistry Essential gene Recombinant DNA biology.protein lcsh:Q Carrier Proteins Sequence Alignment Research Article |
| Zdroj: | PLoS ONE, Vol 10, Iss 11, p e0143770 (2015) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Antifungal drugs acting via new mechanisms of action are urgently needed to combat the increasing numbers of severe fungal infections caused by pathogens such as Candida albicans. The phosphopantetheinyl transferase of Aspergillus fumigatus, encoded by the essential gene pptB, has previously been identified as a potential antifungal target. This study investigated the function of its orthologue in C. albicans, PPT2/C1_09480W by placing one allele under the control of the regulatable MET3 promoter, and deleting the remaining allele. The phenotypes of this conditional null mutant showed that, as in A. fumigatus, the gene PPT2 is essential for growth in C. albicans, thus fulfilling one aspect of an efficient antifungal target. The catalytic activity of Ppt2 as a phosphopantetheinyl transferase and the acyl carrier protein Acp1 as a substrate were demonstrated in a fluorescence transfer assay, using recombinant Ppt2 and Acp1 produced and purified from E.coli. A fluorescence polarisation assay amenable to high-throughput screening was also developed. Therefore we have identified Ppt2 as a broad-spectrum novel antifungal target and developed tools to identify inhibitors as potentially new antifungal compounds. |
| Databáze: | OpenAIRE |
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