Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum
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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Language: | English |
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Frontiers Media
2017-04-06
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Subjects: | Antimicrobial peptides, Antifungal proteins, Protein mapping, Peptide design, Penicillium digitatum, Postharvest pathology, Penicillium chrysogenum, Synergy, |
Online Access: | http://hdl.handle.net/10261/157962 http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100002428 |
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dig-iata-es-10261-1579622021-12-28T15:53:21Z Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum Garrigues, Sandra Gandía Gómez, Mónica Borics, Attila Marx, Florentine Manzanares, Paloma Marcos López, José Francisco Consejo Superior de Investigaciones Científicas (España) Ministerio de Economía y Competitividad (España) European Commission Ministerio de Educación, Cultura y Deporte (España) Austrian Science Fund CSIC - Unidad de Recursos de Información Científica para la Investigación (URICI) Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy 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. This work was funded by grants BIO2012-34381 and BIO2015-68790-C2-1-R from the “Ministerio de Economía y Competitividad” (Spain) (MINECO/FEDER Funds) and P25894-B20 from the Austrian Science Fund (FWF). SG was recipient of a predoctoral scholarship (FPU13/04584) within the FPU program from “Ministerio de Educación, Cultura y Deporte” (MECD, Spain). Support for publication was provided by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). Peer reviewed 2017-12-04T12:10:40Z 2017-12-04T12:10:40Z 2017-04-06 artículo http://purl.org/coar/resource_type/c_6501 Frontiers in Microbiology 8: 592 (2017) http://hdl.handle.net/10261/157962 10.3389/fmicb.2017.00592 1664-302X http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100002428 28428776 en #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-68790-C2-1-R Publisher's version https://doi.org/10.3389/fmicb.2017.00592 Sí open Frontiers Media |
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Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy |
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Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy Garrigues, Sandra Gandía Gómez, Mónica Borics, Attila Marx, Florentine Manzanares, Paloma Marcos López, José Francisco Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
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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. |
author2 |
Consejo Superior de Investigaciones Científicas (España) |
author_facet |
Consejo Superior de Investigaciones Científicas (España) Garrigues, Sandra Gandía Gómez, Mónica Borics, Attila Marx, Florentine Manzanares, Paloma Marcos López, José Francisco |
format |
artículo |
topic_facet |
Antimicrobial peptides Antifungal proteins Protein mapping Peptide design Penicillium digitatum Postharvest pathology Penicillium chrysogenum Synergy |
author |
Garrigues, Sandra Gandía Gómez, Mónica Borics, Attila Marx, Florentine Manzanares, Paloma Marcos López, José Francisco |
author_sort |
Garrigues, Sandra |
title |
Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
title_short |
Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
title_full |
Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
title_fullStr |
Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
title_full_unstemmed |
Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum |
title_sort |
mapping and identification of antifungal peptides in the putative antifungal protein afpb from the filamentous fungus penicillium digitatum |
publisher |
Frontiers Media |
publishDate |
2017-04-06 |
url |
http://hdl.handle.net/10261/157962 http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100002428 |
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