Nucleoside selectivity of <i>Aspergillus fumigatus</i> nucleoside‐diphosphate kinase

Nguyen, Stephanie; Jovcevski, Blagojce; Pukala, Tara L.; Bruning, John B.

doi:10.1111/febs.15607

Cited by 8 publications

(8 citation statements)

References 65 publications

(72 reference statements)

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“…The CYP51 belongs to the cytochrome P450 monooxygenase superfamily and mediates a crucial step of the synthesis of ergosterol, which is a fungal-specific sterol. The NDK catalyzes the reversible exchange of the γ -phosphate between nucleoside triphosphate (NTP) and nucleoside diphosphate (NDP) [ 37 , 38 ]. Among different reported antifungal mechanisms, rutin, and quercetin, found in our plant extracts, seem to inhibit fungal strains via inhibition of CYP51 enzyme, same as that of azole drugs.…”

Section: Discussionmentioning

confidence: 99%

Profiling of Antifungal Activities and In Silico Studies of Natural Polyphenols from Some Plants

et al. 2021

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A worldwide increase in the incidence of fungal infections, emergence of new fungal strains, and antifungal resistance to commercially available antibiotics indicate the need to investigate new treatment options for fungal diseases. Therefore, the interest in exploring the antifungal activity of medicinal plants has now been increased to discover phyto-therapeutics in replacement to conventional antifungal drugs. The study was conducted to explore and identify the mechanism of action of antifungal agents of edible plants, including Cinnamomum zeylanicum, Cinnamomum tamala, Amomum subulatum, Trigonella foenumgraecum, Mentha piperita, Coriandrum sativum, Lactuca sativa, and Brassica oleraceae var. italica. The antifungal potential was assessed via the disc diffusion method and, subsequently, the extracts were assessed for phytochemicals and total antioxidant activity. Potent polyphenols were detected using high-performance liquid chromatography (HPLC) and antifungal mechanism of action was evaluated in silico. Cinnamomum zeylanicum exhibited antifungal activity against all the tested strains while all plant extracts showed antifungal activity against Fusarium solani. Rutin, kaempferol, and quercetin were identified as common polyphenols. In silico studies showed that rutin displayed the greatest affinity with binding pocket of fungal 14-alpha demethylase and nucleoside diphosphokinase with the binding affinity (Kd, −9.4 and −8.9, respectively), as compared to terbinafine. Results indicated that Cinnamomum zeylanicum and Cinnamomum tamala exert their antifungal effect possibly due to kaempferol and rutin, respectively, or possibly by inhibition of nucleoside diphosphokinase (NDK) and 14-alpha demethylase (CYP51), while Amomum subulatum and Trigonella foenum graecum might exhibit antifungal potential due to quercetin. Overall, the study demonstrates that plant-derived products have a high potential to control fungal infections.

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Section: Discussionmentioning

confidence: 99%

Profiling of Antifungal Activities and In Silico Studies of Natural Polyphenols from Some Plants

et al. 2021

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“…The oligomeric state of enolase, which was buffer‐exchanged into 200 mM ammonium acetate (pH 6.8) using an Amicon Ultra‐Centrifugal Filter (10 000 MWCO) at 4°C, was examined using a Synapt G1 HDMS (Waters) using a nanoelectrospray ionization source 24,37 . Enolase (15 μM) was loaded into gold‐coated borosilicate glass capillaries prepared in‐house.…”

Section: Methodsmentioning

confidence: 99%

“…The vector was transformed into E. coli BL21 (λDE3). Methods used for recombinant protein expression, lysis, and purification using nickel‐affinity chromatography have been described previously 24 . Recombinant enolase was purified to >90% homogeneity (assessed by SDS‐PAGE) and buffer‐exchanged into Storage Buffer (50 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 5% v/v glycerol, 1 mM DTT) using size‐exclusion chromatography (HiPrep™ 26/60 Sephacryl® S‐200 HR, GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

“…Methods used for recombinant protein expression, lysis, and purification using nickel-affinity chromatography have been described previously. 24 Recombinant enolase was purified to >90% homogeneity (assessed by SDS-PAGE) and buffer-exchanged into Storage Buffer (50 mM Tris-HCl pH 8.0, 0.5 mM EDTA, 5% v/v glycerol, 1 mM DTT) using size-exclusion chromatography (HiPrep™ 26/60 Sephacryl ® S-200 HR, GE Healthcare). Protein was concentrated to 15.5 mg/ml using an Amicon Ultra-15 centrifugal filter unit (30 kDa MWCO) (Merck Millipore), flash cooled in liquid nitrogen and stored at À80 C. A single crystal was mounted onto a cryo-loop, passed through Paratone-N (Hampton Research) and flash frozen in liquid nitrogen.…”

Section: Enolase-pet15b Expression Vector Construction Protein Expres...mentioning

confidence: 99%

“…The oligomeric state of enolase, which was buffer-exchanged into 200 mM ammonium acetate (pH 6.8) using an Amicon Ultra-Centrifugal Filter (10 000 MWCO) at 4 C, was examined using a Synapt G1 HDMS (Waters) using a nanoelectrospray ionization source. 24,37 Enolase (15 μM) was loaded into gold-coated borosilicate glass capillaries prepared in-house. Key instrument parameters were as follows: capillary voltage (kV): 1.65; sampling cone (V): 50; extraction cone (V): 2.0; trap/transfer collision energy (V/V): 15/10; trap gas (L/h): 5.5; backing gas (mbar): $4.5.…”

Section: Native Mass Spectrometrymentioning

confidence: 99%

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A structural model of the human plasminogen and Aspergillus fumigatus enolase complex

et al. 2022

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The metabolic enzyme, enolase, plays a crucial role in the cytoplasm where it maintains cellular energy production within the process of glycolysis. The main role of enolase in glycolysis is to convert 2-phosphoglycerate to phosphoenolpyruvate; however, enolase can fulfill roles that deviate from this function. In pathogenic bacteria and fungi, enolase is also located on the cell surface where it functions as a virulence factor. Surface-expressed enolase is a receptor for human plasma proteins, including plasminogen, and this interaction facilitates nutrient acquisition and tissue invasion.A novel approach to developing antifungal drugs is to inhibit the formation of this complex. To better understand the structure of enolase and the interactions that may govern complex formation, we have solved the first X-ray crystal structure of enolase from Aspergillus fumigatus (2.0 Å) and have shown that it preferentially adopts a dimeric quaternary structure using native mass spectrometry. Two additional X-ray crystal structures of A. fumigatus enolase bound to the endogenous substrate 2-phosphoglycerate and product phosphoenolpyruvate were determined and kinetic characterization was carried out to better understand the details of its canonical function. From these data, we have produced a model of the A. fumigatus enolase and human plasminogen complex to provide structural insights into the mechanisms of virulence and aid future development of small molecules or peptidomimetics for antifungal drug design.

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Gaining molecular insights towards inhibition of foodborne fungi Aspergillus fumigatus by a food colourant violacein via computational approach

Sindhu,

Bhat,

Sangta

et al. 2024

Sci Rep

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Filamentous Fungal Human Pathogens (FFHPs) such as Aspergillus fumigatus , are growing resistant to currently available antifungal drugs. One possible target, the Nucleoside diphosphate kinase (Ndk) is significant for nucleotide biosynthesis and crucial for fungal metabolism. Violacein, a natural food colorant, was examined for its antifungal effects against Aspergillus fumigatus via computational approach against the Ndk protein. Known and predicted interactions of Ndk with proteins was performed using the STRING application. Molecular docking was performed using Schrodinger Maestro software (V.14.1) under enhanced precision docking, with OPLS4 forcefield. MDS was performed for 500ns under OPLS4 forcefield and the TIP3P solvent system. The geometry optimization for DFT was performed using the Becke 3-parameter exchange functional (B3LYP) method. The Molecular Docking Studies revealed significant interactions with good binding energy between Violacein and Ndk. Subsequent MD Simulations confirmed the stability of Violacein-Ndk complex, compared to the reference ligand-complex, indicating a stable interaction between the protein and violacein. The energy band gap of violacein was found to be 0.072567 eV suggesting its softness with lower kinetic stability and higher chemical reactivity. The results suggest Violacein could potentially disrupt nucleotide metabolism by targeting Ndk, thus demonstrating antifungal activity. However, further experimental validation is required to confirm these computational findings and explore the practical use of Violacein in antifungal treatments.

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Nucleoside selectivity of Aspergillus fumigatus nucleoside‐diphosphate kinase

Cited by 8 publications

References 65 publications

Profiling of Antifungal Activities and In Silico Studies of Natural Polyphenols from Some Plants

Profiling of Antifungal Activities and In Silico Studies of Natural Polyphenols from Some Plants

A structural model of the human plasminogen and Aspergillus fumigatus enolase complex

Gaining molecular insights towards inhibition of foodborne fungi Aspergillus fumigatus by a food colourant violacein via computational approach

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