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The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization

HsAFP1, a plant defensin isolated from coral bells (Heuchera sanguinea), is characterized by broad-spectrum antifungal activity. Previous studies indicated that HsAFP1 binds to specific fungal membrane components, which had hitherto not been identified, and induces mitochondrial dysfunction and cell...

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Autores principales: Cools, Tanne L., Vriens, Kim, Struyfs, Caroline, Verbandt, Sara, Ramada, Marcelo H. S., Brand, Guilherme D., Bloch, Carlos, Koch, Barbara, Traven, Ana, Drijfhout, Jan W., Demuyser, Liesbeth, Kucharíková, Soňa, Van Dijck, Patrick, Spasic, Dragana, Lammertyn, Jeroen, Cammue, Bruno P. A., Thevissen, Karin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5702387/
https://www.ncbi.nlm.nih.gov/pubmed/29209301
http://dx.doi.org/10.3389/fmicb.2017.02295
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author Cools, Tanne L.
Vriens, Kim
Struyfs, Caroline
Verbandt, Sara
Ramada, Marcelo H. S.
Brand, Guilherme D.
Bloch, Carlos
Koch, Barbara
Traven, Ana
Drijfhout, Jan W.
Demuyser, Liesbeth
Kucharíková, Soňa
Van Dijck, Patrick
Spasic, Dragana
Lammertyn, Jeroen
Cammue, Bruno P. A.
Thevissen, Karin
author_facet Cools, Tanne L.
Vriens, Kim
Struyfs, Caroline
Verbandt, Sara
Ramada, Marcelo H. S.
Brand, Guilherme D.
Bloch, Carlos
Koch, Barbara
Traven, Ana
Drijfhout, Jan W.
Demuyser, Liesbeth
Kucharíková, Soňa
Van Dijck, Patrick
Spasic, Dragana
Lammertyn, Jeroen
Cammue, Bruno P. A.
Thevissen, Karin
author_sort Cools, Tanne L.
collection PubMed
description HsAFP1, a plant defensin isolated from coral bells (Heuchera sanguinea), is characterized by broad-spectrum antifungal activity. Previous studies indicated that HsAFP1 binds to specific fungal membrane components, which had hitherto not been identified, and induces mitochondrial dysfunction and cell membrane permeabilization. In this study, we show that HsAFP1 reversibly interacts with the membrane phospholipid phosphatidic acid (PA), which is a precursor for the biosynthesis of other phospholipids, and to a lesser extent with various phosphatidyl inositol phosphates (PtdInsP’s). Moreover, via reverse ELISA assays we identified two basic amino acids in HsAFP1, namely histidine at position 32 and arginine at position 52, as well as the phosphate group in PA as important features enabling this interaction. Using a HsAFP1 variant, lacking both amino acids (HsAFP1[H32A][R52A]), we showed that, as compared to the native peptide, the ability of this variant to bind to PA and PtdInsP’s is reduced (≥74%) and the antifungal activity of the variant is reduced (≥2-fold), highlighting the link between PA/PtdInsP binding and antifungal activity. Using fluorescently labelled HsAFP1 in confocal microscopy and flow cytometry assays, we showed that HsAFP1 accumulates at the cell surface of yeast cells with intact membranes, most notably at the buds and septa. The resulting HsAFP1-induced membrane permeabilization is likely to occur after HsAFP1’s internalization. These data provide novel mechanistic insights in the mode of action of the HsAFP1 plant defensin.
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spelling pubmed-57023872017-12-05 The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization Cools, Tanne L. Vriens, Kim Struyfs, Caroline Verbandt, Sara Ramada, Marcelo H. S. Brand, Guilherme D. Bloch, Carlos Koch, Barbara Traven, Ana Drijfhout, Jan W. Demuyser, Liesbeth Kucharíková, Soňa Van Dijck, Patrick Spasic, Dragana Lammertyn, Jeroen Cammue, Bruno P. A. Thevissen, Karin Front Microbiol Microbiology HsAFP1, a plant defensin isolated from coral bells (Heuchera sanguinea), is characterized by broad-spectrum antifungal activity. Previous studies indicated that HsAFP1 binds to specific fungal membrane components, which had hitherto not been identified, and induces mitochondrial dysfunction and cell membrane permeabilization. In this study, we show that HsAFP1 reversibly interacts with the membrane phospholipid phosphatidic acid (PA), which is a precursor for the biosynthesis of other phospholipids, and to a lesser extent with various phosphatidyl inositol phosphates (PtdInsP’s). Moreover, via reverse ELISA assays we identified two basic amino acids in HsAFP1, namely histidine at position 32 and arginine at position 52, as well as the phosphate group in PA as important features enabling this interaction. Using a HsAFP1 variant, lacking both amino acids (HsAFP1[H32A][R52A]), we showed that, as compared to the native peptide, the ability of this variant to bind to PA and PtdInsP’s is reduced (≥74%) and the antifungal activity of the variant is reduced (≥2-fold), highlighting the link between PA/PtdInsP binding and antifungal activity. Using fluorescently labelled HsAFP1 in confocal microscopy and flow cytometry assays, we showed that HsAFP1 accumulates at the cell surface of yeast cells with intact membranes, most notably at the buds and septa. The resulting HsAFP1-induced membrane permeabilization is likely to occur after HsAFP1’s internalization. These data provide novel mechanistic insights in the mode of action of the HsAFP1 plant defensin. Frontiers Media S.A. 2017-11-21 /pmc/articles/PMC5702387/ /pubmed/29209301 http://dx.doi.org/10.3389/fmicb.2017.02295 Text en Copyright © 2017 Cools, Vriens, Struyfs, Verbandt, Ramada, Brand, Bloch, Koch, Traven, Drijfhout, Demuyser, Kucharíková, Van Dijck, Spasic, Lammertyn, Cammue and Thevissen. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Cools, Tanne L.
Vriens, Kim
Struyfs, Caroline
Verbandt, Sara
Ramada, Marcelo H. S.
Brand, Guilherme D.
Bloch, Carlos
Koch, Barbara
Traven, Ana
Drijfhout, Jan W.
Demuyser, Liesbeth
Kucharíková, Soňa
Van Dijck, Patrick
Spasic, Dragana
Lammertyn, Jeroen
Cammue, Bruno P. A.
Thevissen, Karin
The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title_full The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title_fullStr The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title_full_unstemmed The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title_short The Antifungal Plant Defensin HsAFP1 Is a Phosphatidic Acid-Interacting Peptide Inducing Membrane Permeabilization
title_sort antifungal plant defensin hsafp1 is a phosphatidic acid-interacting peptide inducing membrane permeabilization
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5702387/
https://www.ncbi.nlm.nih.gov/pubmed/29209301
http://dx.doi.org/10.3389/fmicb.2017.02295
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