Genetic and structural validation of Aspergillus fumigatus N-acetylphosphoglucosamine mutase as an antifungal target

Autor: Daan M. F. van Aalten, Michael A. J. Ferguson, Wenxia Fang, Karina Mariño, Ramon Hurtado-Guerrero, Osama Albarbarawi, Cheng Jin, Olawale G. Raimi, Adel F. M. Ibrahim, Ting-Ting Du
Přispěvatelé: Medical Research Council (UK), National Natural Science Foundation of China, Wellcome Trust
Rok vydání: 2013
Předmět:
Models
Molecular
Antifungal Agents
lcsh:Life
lcsh:QR1-502
Drug target
UDP
uridine diphosphate
Crystallography
X-Ray
Nucleotide sugar
Biochemistry
Protein Structure
Secondary
lcsh:Microbiology
Aspergillus fumigatus
purl.org/becyt/ford/1 [https]
RMSD
root mean square deviation
chemistry.chemical_compound
Mutase
Catalytic Domain
Magnesium
CELL WALL
health care economics and organizations
chemistry.chemical_classification
Fru-6P
fructose 6-phosphate
0303 health sciences
AfAGM1
A. fumigatus N-acetylphosphoglucosamine mutase
biology
AGM1
N-acetylphosphoglucosamine mutase
GlcNAc
N-acetylglucosamine
Bioquímica y Biología Molecular
GlcNAc-1P
N-acetylglucosamine-1-phosphate
3. Good health
Phosphotransferases (Phosphomutases)
Cell walls
CIENCIAS NATURALES Y EXACTAS
Inhibitor
MIC
minimum inhibitory concentration
UAP1
UDP–GlcNAc pyrophosphorylase
Biophysics
Microbial Sensitivity Tests
MM
minimal medium
S2
Microbiology
Fungal Proteins
Ciencias Biológicas
Cell wall
Inhibitory Concentration 50
03 medical and health sciences
Chitin
G6PDH
glucose-6-phosphate dehydrogenase
Glycosyltransferase
GFA1
glutamine: Fru-6P amidotransferase
Humans
purl.org/becyt/ford/1.6 [https]
GlcN-6P
glucosamine 6-phosphate
Molecular Biology
GNA1
GlcN-6P acetyltransferase
030304 developmental biology
Original Paper
Microbial Viability
030306 microbiology
Cell Biology
biology.organism_classification
Kinetics
lcsh:QH501-531
Uridine diphosphate
Enzyme
chemistry
CaAGM1
Candida albicans AGM1
biology.protein
Protein structure
cell wall
IA
invasive aspergillosis
Zdroj: Scopus-Elsevier
Digital.CSIC: Repositorio Institucional del CSIC
Consejo Superior de Investigaciones Científicas (CSIC)
Bioscience Reports, Vol 33, Iss 5, p e00063 (2013)
Bioscience Reports
Digital.CSIC. Repositorio Institucional del CSIC
instname
CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
ISSN: 0144-8463
3103-0025
Popis: 11 pags, 5 figs, 3 tabs
Synopsis Aspergillus fumigatus is the causative agent of IA (invasive aspergillosis) in immunocompromised patients. It possesses a cell wall composed of chitin, glucan and galactomannan, polymeric carbohydrates synthesized by processive glycosyltransferases from intracellular sugar nucleotide donors. Here we demonstrate that A. fumigatus possesses an active AfAGM1 (A. fumigatus N-acetylphosphoglucosamine mutase), a key enzyme in the biosynthesis of UDP (uridine diphosphate)-GlcNAc (N-acetylglucosamine), the nucleotide sugar donor for chitin synthesis. A conditional agm1 mutant revealed the gene to be essential. Reduced expression of agm1 resulted in retarded cell growth and altered cell wall ultrastructure and composition. The crystal structure of AfAGM1 revealed an amino acid change in the active site compared with the human enzyme, which could be exploitable in the design of selective inhibitors. AfAGM1 inhibitors were discovered by high-throughput screening, inhibiting the enzyme with IC50s in the low μM range. Together, these data provide a platform for the future development of AfAGM1 inhibitors with antifungal activity. © 2013 The Author(s).
This work was supported by the Medical Research Council [Programme Grant number G0900138] and the National Natural Science Foundation of China [grant number 31030025] (to C.J.). D.v.A. is funded by a Wellcome Trust Senior Research Fellowship [grant number WT087590MA]. M.A.J.F. and K.M. were supported by a Wellcome Trust Programme Grant [grant number 085622].
Databáze: OpenAIRE
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