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Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)

The PA1b (Pea Albumin 1, subunit b) peptide is an entomotoxin extract from Legume seeds with lethal activity on several insect pests, such as mosquitoes, some aphids and cereal weevils. This 37 amino-acid cysteine-rich peptide has been, until now, obtained by biochemical purification or chemical syn...

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Autores principales: Eyraud, Vanessa, Karaki, Lamis, Rahioui, Isabelle, Sivignon, Catherine, Da Silva, Pedro, Rahbé, Yvan, Royer, Corinne, Gressent, Frédéric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3859497/
https://www.ncbi.nlm.nih.gov/pubmed/24349099
http://dx.doi.org/10.1371/journal.pone.0081619
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author Eyraud, Vanessa
Karaki, Lamis
Rahioui, Isabelle
Sivignon, Catherine
Da Silva, Pedro
Rahbé, Yvan
Royer, Corinne
Gressent, Frédéric
author_facet Eyraud, Vanessa
Karaki, Lamis
Rahioui, Isabelle
Sivignon, Catherine
Da Silva, Pedro
Rahbé, Yvan
Royer, Corinne
Gressent, Frédéric
author_sort Eyraud, Vanessa
collection PubMed
description The PA1b (Pea Albumin 1, subunit b) peptide is an entomotoxin extract from Legume seeds with lethal activity on several insect pests, such as mosquitoes, some aphids and cereal weevils. This 37 amino-acid cysteine-rich peptide has been, until now, obtained by biochemical purification or chemical synthesis. In this paper, we present our results for the transient production of the peptide in Nicotiana benthamiana by agro-infiltration, with a yield of about 35 µg/g of fresh leaves and maximum production 8 days after infiltration. PA1b is part of the PA1 gene which, after post-translational modifications, encodes two peptides (PA1b and PA1a). We show that transforming tobacco with the PA1b cDNA alone does not result in production of the toxin and, in fact, the entire cDNA is necessary, raising the question of the role of PA1a. We constructed a PA1-cassette, allowing for the quick “cut/paste” of different PA1b mutants within a conserved PA1 cDNA. This cassette enabled us to produce the six isoforms of PA1b which exist in pea seeds. Biological tests revealed that all the isoforms display similar activity, with the exception of one which is inactive. The lack of activity in this isoform led us to conclude that the amphiphilic nature of the peptide is necessary for activity. The possible applications of this expression system for other cysteine-rich biomolecules are discussed.
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spelling pubmed-38594972013-12-13 Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b) Eyraud, Vanessa Karaki, Lamis Rahioui, Isabelle Sivignon, Catherine Da Silva, Pedro Rahbé, Yvan Royer, Corinne Gressent, Frédéric PLoS One Research Article The PA1b (Pea Albumin 1, subunit b) peptide is an entomotoxin extract from Legume seeds with lethal activity on several insect pests, such as mosquitoes, some aphids and cereal weevils. This 37 amino-acid cysteine-rich peptide has been, until now, obtained by biochemical purification or chemical synthesis. In this paper, we present our results for the transient production of the peptide in Nicotiana benthamiana by agro-infiltration, with a yield of about 35 µg/g of fresh leaves and maximum production 8 days after infiltration. PA1b is part of the PA1 gene which, after post-translational modifications, encodes two peptides (PA1b and PA1a). We show that transforming tobacco with the PA1b cDNA alone does not result in production of the toxin and, in fact, the entire cDNA is necessary, raising the question of the role of PA1a. We constructed a PA1-cassette, allowing for the quick “cut/paste” of different PA1b mutants within a conserved PA1 cDNA. This cassette enabled us to produce the six isoforms of PA1b which exist in pea seeds. Biological tests revealed that all the isoforms display similar activity, with the exception of one which is inactive. The lack of activity in this isoform led us to conclude that the amphiphilic nature of the peptide is necessary for activity. The possible applications of this expression system for other cysteine-rich biomolecules are discussed. Public Library of Science 2013-12-11 /pmc/articles/PMC3859497/ /pubmed/24349099 http://dx.doi.org/10.1371/journal.pone.0081619 Text en © 2013 Eyraud et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Eyraud, Vanessa
Karaki, Lamis
Rahioui, Isabelle
Sivignon, Catherine
Da Silva, Pedro
Rahbé, Yvan
Royer, Corinne
Gressent, Frédéric
Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title_full Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title_fullStr Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title_full_unstemmed Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title_short Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b)
title_sort expression and biological activity of the cystine knot bioinsecticide pa1b (pea albumin 1 subunit b)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3859497/
https://www.ncbi.nlm.nih.gov/pubmed/24349099
http://dx.doi.org/10.1371/journal.pone.0081619
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