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

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 Neosart...

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Autores principales: Váradi, Györgyi, Kele, Zoltán, Czajlik, András, Borics, Attila, Bende, Gábor, Papp, Csaba, Rákhely, Gábor, Tóth, Gábor K., Batta, Gyula, Galgóczy, László
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10273333/
https://www.ncbi.nlm.nih.gov/pubmed/37272210
http://dx.doi.org/10.1002/pro.4692
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author Váradi, Györgyi
Kele, Zoltán
Czajlik, András
Borics, Attila
Bende, Gábor
Papp, Csaba
Rákhely, Gábor
Tóth, Gábor K.
Batta, Gyula
Galgóczy, László
author_facet Váradi, Györgyi
Kele, Zoltán
Czajlik, András
Borics, Attila
Bende, Gábor
Papp, Csaba
Rákhely, Gábor
Tóth, Gábor K.
Batta, Gyula
Galgóczy, László
author_sort Váradi, Györgyi
collection PubMed
description 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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spelling pubmed-102733332023-07-01 Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2 Váradi, Györgyi Kele, Zoltán Czajlik, András Borics, Attila Bende, Gábor Papp, Csaba Rákhely, Gábor Tóth, Gábor K. Batta, Gyula Galgóczy, László Protein Sci Articles 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. John Wiley & Sons, Inc. 2023-07-01 /pmc/articles/PMC10273333/ /pubmed/37272210 http://dx.doi.org/10.1002/pro.4692 Text en © 2023 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Váradi, Györgyi
Kele, Zoltán
Czajlik, András
Borics, Attila
Bende, Gábor
Papp, Csaba
Rákhely, Gábor
Tóth, Gábor K.
Batta, Gyula
Galgóczy, László
Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title_full Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title_fullStr Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title_full_unstemmed Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title_short Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2
title_sort hard nut to crack: solving the disulfide linkage pattern of the neosartorya (aspergillus) fischeri antifungal protein 2
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10273333/
https://www.ncbi.nlm.nih.gov/pubmed/37272210
http://dx.doi.org/10.1002/pro.4692
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