Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure

Bacillus thuringiensis, a well-known insecticidal bacterium, produces several insecticidal proteins, including cytolytic (Cyt) proteins. Cyt proteins bind directly to the lipid membrane and form large protein complexes. In addition to the protein ladder bands, information on the oligomeric structure...

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Main Authors: Chontida Tangsongcharoen, Jose L. Toca-Herrera, Boonhiang Promdonkoy, Kanokporn Srisucharitpanit, Sudarat Tharad
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Toxicon: X
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Online Access:http://www.sciencedirect.com/science/article/pii/S2590171025000074
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author Chontida Tangsongcharoen
Jose L. Toca-Herrera
Boonhiang Promdonkoy
Kanokporn Srisucharitpanit
Sudarat Tharad
author_facet Chontida Tangsongcharoen
Jose L. Toca-Herrera
Boonhiang Promdonkoy
Kanokporn Srisucharitpanit
Sudarat Tharad
author_sort Chontida Tangsongcharoen
collection DOAJ
description Bacillus thuringiensis, a well-known insecticidal bacterium, produces several insecticidal proteins, including cytolytic (Cyt) proteins. Cyt proteins bind directly to the lipid membrane and form large protein complexes. In addition to the protein ladder bands, information on the oligomeric structure in lipid membranes is necessary to understand the mechanism of Cyt proteins on target cells. In this work, we have investigated the oligomeric Cyt2Aa2 complex with synthetic lipid and with erythrocyte membranes. When the activated Cyt2Aa2 protein was incubated with these lipid membranes, the protein ladder pattern relevant to hemolytic activity was detected in SDS-PAGE. Moreover, AFM topographic images revealed a fusilli-like structure and a ring-like structure for synthetic POPC and POPC/Chol, respectively. Furthermore, TEM micrographs provided an additional information on the oligomeric structure of Cyt2Aa2 in erythrocytes. Cyt2Aa2 appears to oligomerise/aggregate into mixed structures between the filamentous structure and small protein complexes in erythrocytes. In addition, a nanopore was found to be a substructure of the filamentous structure. These results strengthen the understanding of Cyt2Aa2 behavior in these two membrane systems, the fusilli and ring-like structures, depending on the type of lipid membrane. Furthermore, the structure of Cyt2Aa2 in insect target membranes remains to be investigated.
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spelling doaj-art-8ea88f30bcde44368523c5ed323463422025-08-20T02:05:07ZengElsevierToxicon: X2590-17102025-06-012610022010.1016/j.toxcx.2025.100220Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structureChontida Tangsongcharoen0Jose L. Toca-Herrera1Boonhiang Promdonkoy2Kanokporn Srisucharitpanit3Sudarat Tharad4Faculty of Allied Health Sciences, Burapha University, Chonburi, 20131, ThailandInstitut für Biophysik, Department für Bionanowissenschaften, Universität für Bodenkultur Wien (BOKU), Vienna, 1190, AustriaNational Center for Genetic Engineering and Biotechnology, National Science and Technology, Development Agency, Pathumthani, 12120, ThailandFaculty of Allied Health Sciences, Burapha University, Chonburi, 20131, ThailandDepartment of Biology, Faculty of Science, Burapha University, Chonburi, 20131, Thailand; Corresponding author.Bacillus thuringiensis, a well-known insecticidal bacterium, produces several insecticidal proteins, including cytolytic (Cyt) proteins. Cyt proteins bind directly to the lipid membrane and form large protein complexes. In addition to the protein ladder bands, information on the oligomeric structure in lipid membranes is necessary to understand the mechanism of Cyt proteins on target cells. In this work, we have investigated the oligomeric Cyt2Aa2 complex with synthetic lipid and with erythrocyte membranes. When the activated Cyt2Aa2 protein was incubated with these lipid membranes, the protein ladder pattern relevant to hemolytic activity was detected in SDS-PAGE. Moreover, AFM topographic images revealed a fusilli-like structure and a ring-like structure for synthetic POPC and POPC/Chol, respectively. Furthermore, TEM micrographs provided an additional information on the oligomeric structure of Cyt2Aa2 in erythrocytes. Cyt2Aa2 appears to oligomerise/aggregate into mixed structures between the filamentous structure and small protein complexes in erythrocytes. In addition, a nanopore was found to be a substructure of the filamentous structure. These results strengthen the understanding of Cyt2Aa2 behavior in these two membrane systems, the fusilli and ring-like structures, depending on the type of lipid membrane. Furthermore, the structure of Cyt2Aa2 in insect target membranes remains to be investigated.http://www.sciencedirect.com/science/article/pii/S2590171025000074Bacillus thuringiensisCytolytic proteinCyt2Aa2 proteinOligomer assemblyFusilli-like structureThread-like filament structure
spellingShingle Chontida Tangsongcharoen
Jose L. Toca-Herrera
Boonhiang Promdonkoy
Kanokporn Srisucharitpanit
Sudarat Tharad
Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
Toxicon: X
Bacillus thuringiensis
Cytolytic protein
Cyt2Aa2 protein
Oligomer assembly
Fusilli-like structure
Thread-like filament structure
title Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
title_full Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
title_fullStr Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
title_full_unstemmed Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
title_short Oligomer assembly of Bacillus thuringiensis Cyt2Aa2 on lipid membranes reveals a thread-like structure
title_sort oligomer assembly of bacillus thuringiensis cyt2aa2 on lipid membranes reveals a thread like structure
topic Bacillus thuringiensis
Cytolytic protein
Cyt2Aa2 protein
Oligomer assembly
Fusilli-like structure
Thread-like filament structure
url http://www.sciencedirect.com/science/article/pii/S2590171025000074
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