Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.

<h4>Background</h4>Allium sativum leaf agglutinin (ASAL) is a 25-kDa homodimeric, insecticidal, mannose binding lectin whose subunits are assembled by the C-terminal exchange process. An attempt was made to convert dimeric ASAL into a monomeric form to correlate the relevance of quaterna...

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Autores principales:	Nilanjana Banerjee, Subhadipa Sengupta, Amit Roy, Prithwi Ghosh, Kalipada Das, Sampa Das
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fd47e2dfa5bc4c6e8f39e6a1495a4cc2
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spelling	oai:doaj.org-article:fd47e2dfa5bc4c6e8f39e6a1495a4cc22021-11-18T06:56:06ZFunctional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.1932-620310.1371/journal.pone.0018593https://doaj.org/article/fd47e2dfa5bc4c6e8f39e6a1495a4cc22011-04-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21490929/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Allium sativum leaf agglutinin (ASAL) is a 25-kDa homodimeric, insecticidal, mannose binding lectin whose subunits are assembled by the C-terminal exchange process. An attempt was made to convert dimeric ASAL into a monomeric form to correlate the relevance of quaternary association of subunits and their functional specificity. Using SWISS-MODEL program a stable monomer was designed by altering five amino acid residues near the C-terminus of ASAL.<h4>Methodology/principal findings</h4>By introduction of 5 site-specific mutations (-DNSNN-), a β turn was incorporated between the 11(th) and 12(th) β strands of subunits of ASAL, resulting in a stable monomeric mutant ASAL (mASAL). mASAL was cloned and subsequently purified from a pMAL-c2X system. CD spectroscopic analysis confirmed the conservation of secondary structure in mASAL. Mannose binding assay confirmed that molecular mannose binds efficiently to both mASAL and ASAL. In contrast to ASAL, the hemagglutination activity of purified mASAL against rabbit erythrocytes was lost. An artificial diet bioassay of Lipaphis erysimi with mASAL displayed an insignificant level of insecticidal activity compared to ASAL. Fascinatingly, mASAL exhibited strong antifungal activity against the pathogenic fungi Fusarium oxysporum, Rhizoctonia solani and Alternaria brassicicola in a disc diffusion assay. A propidium iodide uptake assay suggested that the inhibitory activity of mASAL might be associated with the alteration of the membrane permeability of the fungus. Furthermore, a ligand blot assay of the membrane subproteome of R. solani with mASAL detected a glycoprotein receptor having interaction with mASAL.<h4>Conclusions/significance</h4>Conversion of ASAL into a stable monomer resulted in antifungal activity. From an evolutionary aspect, these data implied that variable quaternary organization of lectins might be the outcome of defense-related adaptations to diverse situations in plants. Incorporation of mASAL into agronomically-important crops could be an alternative method to protect them from dramatic yield losses from pathogenic fungi in an effective manner.Nilanjana BanerjeeSubhadipa SenguptaAmit RoyPrithwi GhoshKalipada DasSampa DasPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 4, p e18593 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nilanjana Banerjee Subhadipa Sengupta Amit Roy Prithwi Ghosh Kalipada Das Sampa Das Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
description	<h4>Background</h4>Allium sativum leaf agglutinin (ASAL) is a 25-kDa homodimeric, insecticidal, mannose binding lectin whose subunits are assembled by the C-terminal exchange process. An attempt was made to convert dimeric ASAL into a monomeric form to correlate the relevance of quaternary association of subunits and their functional specificity. Using SWISS-MODEL program a stable monomer was designed by altering five amino acid residues near the C-terminus of ASAL.<h4>Methodology/principal findings</h4>By introduction of 5 site-specific mutations (-DNSNN-), a β turn was incorporated between the 11(th) and 12(th) β strands of subunits of ASAL, resulting in a stable monomeric mutant ASAL (mASAL). mASAL was cloned and subsequently purified from a pMAL-c2X system. CD spectroscopic analysis confirmed the conservation of secondary structure in mASAL. Mannose binding assay confirmed that molecular mannose binds efficiently to both mASAL and ASAL. In contrast to ASAL, the hemagglutination activity of purified mASAL against rabbit erythrocytes was lost. An artificial diet bioassay of Lipaphis erysimi with mASAL displayed an insignificant level of insecticidal activity compared to ASAL. Fascinatingly, mASAL exhibited strong antifungal activity against the pathogenic fungi Fusarium oxysporum, Rhizoctonia solani and Alternaria brassicicola in a disc diffusion assay. A propidium iodide uptake assay suggested that the inhibitory activity of mASAL might be associated with the alteration of the membrane permeability of the fungus. Furthermore, a ligand blot assay of the membrane subproteome of R. solani with mASAL detected a glycoprotein receptor having interaction with mASAL.<h4>Conclusions/significance</h4>Conversion of ASAL into a stable monomer resulted in antifungal activity. From an evolutionary aspect, these data implied that variable quaternary organization of lectins might be the outcome of defense-related adaptations to diverse situations in plants. Incorporation of mASAL into agronomically-important crops could be an alternative method to protect them from dramatic yield losses from pathogenic fungi in an effective manner.
format	article
author	Nilanjana Banerjee Subhadipa Sengupta Amit Roy Prithwi Ghosh Kalipada Das Sampa Das
author_facet	Nilanjana Banerjee Subhadipa Sengupta Amit Roy Prithwi Ghosh Kalipada Das Sampa Das
author_sort	Nilanjana Banerjee
title	Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
title_short	Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
title_full	Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
title_fullStr	Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
title_full_unstemmed	Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
title_sort	functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/fd47e2dfa5bc4c6e8f39e6a1495a4cc2
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Functional alteration of a dimeric insecticidal lectin to a monomeric antifungal protein correlated to its oligomeric status.

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