NFAP2, a novel cysteine-rich anti-yeast protein from Neosartorya fischeri NRRL 181: isolation and characterization

Tóth, László; Kele, Zoltán; Borics, Attila; Nagy, László G.; Váradi, Györgyi; Virágh, Máté; Takó, Miklós; Vágvölgyi, Csaba; Galgóczy, László

doi:10.1186/s13568-016-0250-8

Cited by 44 publications

(68 citation statements)

References 41 publications

(63 reference statements)

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“…A significant diversity of AFP-like genes and proteins are widespread in Ascomycetes, with genomes that encode up to three different sequence-related AFPs (Galgóczy et al, 2013; Garrigues et al, 2016; Tóth et al, 2016), providing a rich source of potentially divergent antifungals. Bioinformatic and phylogenetic analyses suggested the classification of the different AFP-like sequences in at least three different classes: A, B, and C (Garrigues et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum

et al. 2017

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Antifungal proteins (AFPs) from Ascomycetes are small cysteine-rich proteins that are abundantly secreted and show antifungal activity against non-producer fungi. A gene coding for a class B AFP (AfpB) was previously identified in the genome of the plant pathogen Penicillium digitatum. However, previous attempts to detect the AfpB protein were not successful despite the high expression of the corresponding afpB gene. In this work, the structure of the putative AfpB was modeled. Based on this model, four synthetic cysteine-containing peptides, PAF109, PAF112, PAF118, and PAF119, were designed and their antimicrobial activity was tested and characterized. PAF109 that corresponds to the γ-core motif present in defensin-like antimicrobial proteins did not show antimicrobial activity. On the contrary, PAF112 and PAF118, which are cationic peptides derived from two surface-exposed loops in AfpB, showed moderate antifungal activity against P. digitatum and other filamentous fungi. It was also confirmed that cyclization through a disulfide bridge prevented peptide degradation. PAF116, which is a peptide analogous to PAF112 but derived from the Penicillium chrysogenum antifungal protein PAF, showed activity against P. digitatum similar to PAF112, but was less active than the native PAF protein. The two AfpB-derived antifungal peptides PAF112 and PAF118 showed positive synergistic interaction when combined against P. digitatum. Furthermore, the synthetic hexapeptide PAF26 previously described in our laboratory also exhibited synergistic interaction with the peptides PAF112, PAF118, and PAF116, as well as with the PAF protein. This study is an important contribution to the mapping of antifungal motifs within the AfpB and other AFPs, and opens up new strategies for the rational design and application of antifungal peptides and proteins.

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Section: Introductionmentioning

confidence: 99%

Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum

et al. 2017

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“…PAF and PAFB (PAF‐clade proteins) from P. chrysogenum ; or to different clades e. g . NFAP (PAF‐clade protein) and NFAP2 (NFAP2‐clade protein) from N. fischeri . The crAFPs are mainly effective against filamentous fungi, but potent anti‐yeast activity has been reported recently only for some representatives belonging to the PAF‐ (AnAFP, FPAP, PAF, PAFB), BP‐(BP), and NFAP2‐clade (NFAP2), respectively ,.…”

Section: Anti‐candida Proteins From Filamentous Ascomycetesmentioning

confidence: 99%

“…Electronic circular dichroism spectroscopy of PAF, PAFB, NFAP2 and structural prediction of AnAFP and FPAP (in this review) revealed that these crAFPs have a common β‐pleated conformation due to the presence of numerous β‐strands in their secondary structure (Figure ) ,,…”

Section: Anti‐candida Proteins From Filamentous Ascomycetesmentioning

confidence: 99%

“…Generally, it is typical for all members of this protein group that high ionic strength media decrease the in vitro antifungal efficacy ,. In contrast to NFAP2, all anti‐yeast crAFPs tested so far inhibit the growth of filamentous fungi,,, but were found inactive against yeasts in standard HCCS test medium, for example, the P. chrysogenum antifungal proteins PAF and PAFB show very low activity against different Candida species in RPMI 1640 (unpublished results). One possible explanation for this phenomenon is provided by the mechanism of action of antifungal plant defensins.…”

Section: Anti‐candida Activity Of Crafpsmentioning

confidence: 99%

“…The anti‐ Candida mechanisms of crAFPs are not fully understood, and available data exist only for NFAP2,, PAF,, and PAFB . NFAP2 causes prompt plasma membrane disruption in C. albicans indicating a fast and fungicidal mechanism of action even in high cationic medium under clinical susceptibility test conditions ,. The fungicidal mechanism reduces the risk of resistance development, and further underlines the antifungal potential of NFAP2 under in vivo conditions.…”

Section: Anti‐candida Activity Of Crafpsmentioning

confidence: 99%

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Cysteine‐Rich Antifungal Proteins from Filamentous Fungi are Promising Bioactive Natural Compounds in Anti‐Candida Therapy

Galgóczy

Yap

Marx

2019

Israel Journal of Chemistry

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The emerging number of life‐threatening invasive fungal infections caused by drug‐resistant Candida strains urges the need for the development and application of fundamentally new and safe antifungal strategies in the clinical treatment. Recent studies demonstrated that the extracellular cysteine‐rich and cationic antifungal proteins (crAFPs) originating from filamentous fungi, and de novo designed synthetic peptide derivatives of these crAFPs provide a feasible basis for this approach. This mini‐review focuses on the global challenges of the anti‐Canidia therapy and on the crAFPs as potential drug candidates to overcome existing problems. The advantages and limitations in the use of crAFPs and peptide derivatives compared to those of conventional antifungal drugs will also be critically discussed.

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Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2

Váradi,

Kele,

Czajlik

et al. 2023

Protein Science

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As a consequence of the fast resistance spreading, a limited number of drugs are available to treat fungal infections. Therefore, there is an urgent need to develop new antifungal treatment strategies. The features of a disulfide bond‐stabilized antifungal protein, NFAP2 secreted by the mold Neosartorya (Aspergillus) fischeri render it to be a promising template for future protein‐based antifungal drug design, which requires knowledge about the native disulfide linkage pattern as it is one of the prerequisites for biological activity. However, in the lack of tryptic and chymotryptic proteolytic sites in the ACNCPNNCK sequence, the determination of the disulfide linkage pattern of NFAP2 is not easy with traditional mass spectrometry‐based methods. According to in silico predictions working with a preliminary nuclear magnetic resonance (NMR) solution structure, two disulfide isomers of NFAP2 (abbacc and abbcac) were possible. Both were chemically synthesized; and comparative reversed‐phase high‐performance liquid chromatography, electronic circular dichroism and NMR spectroscopy analyses, and antifungal susceptibility and efficacy tests indicated that the abbcac is the native pattern. This knowledge allowed rational modification of NAFP2 to improve the antifungal efficacy and spectrum through the modulation of the evolutionarily conserved γ‐core region, which is responsible for the activity of several antimicrobial peptides. Disruption of the steric structure of NFAP2 upon γ‐core modification led to the conclusions that this motif may affect the formation of the biologically active three‐dimensional structure, and that the γ‐core modulation is not an efficient tool to improve the antifungal efficacy or to change the antifungal spectrum of NFAP2.

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NFAP2, a novel cysteine-rich anti-yeast protein from Neosartorya fischeri NRRL 181: isolation and characterization

Cited by 44 publications

References 41 publications

Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum

Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum

Cysteine‐Rich Antifungal Proteins from Filamentous Fungi are Promising Bioactive Natural Compounds in Anti‐Candida Therapy

Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2

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