Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity

Abstract The recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. The structurally very similar cysteine-rich antifungal proteins from ascomycetes provide a feasible basis for designing new antifungal molecules. The main st...

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Autores principales:	László Galgóczy, Attila Borics, Máté Virágh, Hargita Ficze, Györgyi Váradi, Zoltán Kele, Florentine Marx
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/802f6340ad9b479098fbe2d69989e9e7
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spelling	oai:doaj.org-article:802f6340ad9b479098fbe2d69989e9e72021-12-02T12:32:57ZStructural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity10.1038/s41598-017-02234-w2045-2322https://doaj.org/article/802f6340ad9b479098fbe2d69989e9e72017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02234-whttps://doaj.org/toc/2045-2322Abstract The recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. The structurally very similar cysteine-rich antifungal proteins from ascomycetes provide a feasible basis for designing new antifungal molecules. The main structural elements responsible for folding, stability and antifungal activity are not fully understood, although this is an essential prerequisite for rational protein design. In this study, we used the Neosartorya fischeri antifungal protein (NFAP) to investigate the role of the disulphide bridges, the hydrophobic core, and the N-terminal amino acids in the formation of a highly stable, folded, and antifungal active protein. NFAP and its mutants carrying cysteine deletion (NFAPΔC), hydrophobic core deletion (NFAPΔh), and N-terminal amino acids exchanges (NFAPΔN) were produced in Pichia pastoris. The recombinant NFAP showed the same features in structure, folding, stability and activity as the native protein. The data acquired with mass spectrometry, structural analyses and antifungal activity assays of NFAP and its mutants proved the importance of the disulphide bonding, the hydrophobic core and the correct N-terminus for folding, stability and full antifungal function. Our findings provide further support to the comprehensive understanding of the structure-function relationship in members of this protein group.László GalgóczyAttila BoricsMáté VirághHargita FiczeGyörgyi VáradiZoltán KeleFlorentine MarxNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q László Galgóczy Attila Borics Máté Virágh Hargita Ficze Györgyi Váradi Zoltán Kele Florentine Marx Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
description	Abstract The recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. The structurally very similar cysteine-rich antifungal proteins from ascomycetes provide a feasible basis for designing new antifungal molecules. The main structural elements responsible for folding, stability and antifungal activity are not fully understood, although this is an essential prerequisite for rational protein design. In this study, we used the Neosartorya fischeri antifungal protein (NFAP) to investigate the role of the disulphide bridges, the hydrophobic core, and the N-terminal amino acids in the formation of a highly stable, folded, and antifungal active protein. NFAP and its mutants carrying cysteine deletion (NFAPΔC), hydrophobic core deletion (NFAPΔh), and N-terminal amino acids exchanges (NFAPΔN) were produced in Pichia pastoris. The recombinant NFAP showed the same features in structure, folding, stability and activity as the native protein. The data acquired with mass spectrometry, structural analyses and antifungal activity assays of NFAP and its mutants proved the importance of the disulphide bonding, the hydrophobic core and the correct N-terminus for folding, stability and full antifungal function. Our findings provide further support to the comprehensive understanding of the structure-function relationship in members of this protein group.
format	article
author	László Galgóczy Attila Borics Máté Virágh Hargita Ficze Györgyi Váradi Zoltán Kele Florentine Marx
author_facet	László Galgóczy Attila Borics Máté Virágh Hargita Ficze Györgyi Váradi Zoltán Kele Florentine Marx
author_sort	László Galgóczy
title	Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
title_short	Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
title_full	Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
title_fullStr	Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
title_full_unstemmed	Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity
title_sort	structural determinants of neosartorya fischeri antifungal protein (nfap) for folding, stability and antifungal activity
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/802f6340ad9b479098fbe2d69989e9e7
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Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity

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