Structure of the Lysinibacillus sphaericus Tpp49Aa1 pesticidal protein elucidated from natural crystals using MHz-SFX
Year: 2023
Type: article
Abstract: The Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito Culex quinquefasciatus and have potential use in biocontrol. Given that proteins with sequence homology to the individual proteins can have activity alone against other insect species, the structure of Tpp49Aa1 was solved in order to understand this protein more fully and inform the design of improved biopesticides. Tpp49Aa1 is naturally expressed as a crystalline inclusion within the host bacterium, and MHz serial femtosecond crystallography using the novel nanofocus option at an X-ray free electron laser allowed rapid and high-quality data collection to determine the structure of Tpp49Aa1 at 1.62 Å resolution. This revealed the packing of Tpp49Aa1 within these natural nanocrystals as a homodimer with a large intermolecular interface. Complementary experiments conducted at varied pH also enabled investigation of the early structural events leading up to the dissolution of natural Tpp49Aa1 crystals—a crucial step in its mechanism of action. To better understand the cooperation between the two proteins, assays were performed on a range of different mosquito cell lines using both individual proteins and mixtures of the two. Finally, bioassays demonstrated Tpp49Aa1/Cry48Aa1 susceptibility of Anopheles stephensi , Aedes albopictus, and Culex tarsalis larvae—substantially increasing the potential use of this binary toxin in mosquito control.
Authors: Lainey J. Williamson, Marina Galchenkova, Hannah L. Best, Richard J. Bean, Anna Munke, Salah Awel, Gisel Pena, Juraj Knoska, Robin Schubert, Katerina Dörner, Hyun-Woo Park, Dennis K. Bideshi, Alessandra Henkel, Viviane Kremling, Bjarne Klopprogge, Emyr Lloyd-Evans, Mark T. Young, Joana Valerio, Marco Kloos, Marcin Sikorski, Grant Mills, Johan Bielecki, Henry Kirkwood, Chan Kim, Raphael de Wijn, Kristina Lorenzen, Paul Lourdu Xavier, Aida Rahmani Mashhour, Luca Gelisio, Oleksandr Yefanov, Adrian P. Mancuso, Brian A. Federici, Henry N. Chapman, Neil Crickmore, Pierre J. Rizkallah, Colin Berry, Dominik Oberthür
Institutions: Cardiff University, Deutsches Elektronen-Synchrotron DESY, Center for Free-Electron Laser Science, European X-Ray Free-Electron Laser, California Baptist University, Max Planck Institute for the Structure and Dynamics of Matter, La Trobe University, University of California, Riverside, Institute for Integrative Genome Biology, Universität Hamburg, Hamburg Centre for Ultrafast Imaging, University of Sussex
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Funders: Joachim Herz Stiftung, UK Research and Innovation, Llywodraeth Cymru, Centers for Disease Control and Prevention, Directorate for Biological Sciences, University of California, Irvine, Biotechnology and Biological Sciences Research Council, Deutsches Elektronen-Synchrotron, European Regional Development Fund
Awards: Characterising a critical role for novel glycosphingolipids in the mechanism of action of insecticidal Toxin_10 proteins, G336278089, G3524225081, Molecular analysis of the interaction of Bin pesticidal protein with its receptor, South West Biosciences: A Doctoral Training Programme for Bioscience students at Bristol, Bath, Cardiff, Exeter and Rothamsted Research, G633442007, PACIFIC SOUTHWEST REGIONAL CENTER OF EXCELLENCE FOR VECTOR-BORNE DISEASE RESEARCH AT THE UNIVERSITY OF CALIFORNIA
OpenAlex ID: W4389083124