Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade
Year: 2020
Type: article
Abstract: Cyt1Aa is the one of four crystalline protoxins produced by mosquitocidal bacterium Bacillus thuringiensis israelensis (Bti) that has been shown to delay the evolution of insect resistance in the field. Limiting our understanding of Bti efficacy and the path to improved toxicity and spectrum has been ignorance of how Cyt1Aa crystallizes in vivo and of its mechanism of toxicity. Here, we use serial femtosecond crystallography to determine the Cyt1Aa protoxin structure from sub-micron-sized crystals produced in Bti. Structures determined under various pH/redox conditions illuminate the role played by previously uncharacterized disulfide-bridge and domain-swapped interfaces from crystal formation in Bti to dissolution in the larval mosquito midgut. Biochemical, toxicological and biophysical methods enable the deconvolution of key steps in the Cyt1Aa bioactivation cascade. We additionally show that the size, shape, production yield, pH sensitivity and toxicity of Cyt1Aa crystals grown in Bti can be controlled by single atom substitution.
Source: Nature Communications
Authors: Guillaume Tetreau, Anne-Sophie Banneville, Elena A. Andreeva, Aaron S. Brewster, Mark S. Hunter, Raymond G. Sierra, Jean-Marie Teulon, Iris D. Young, Niamh Burke, Tilman A. Grünewald, Joël Beaudouin, Irina Snigireva, Maria Teresa Fernandez-Luna, Alister Burt, Hyun-Woo Park, Luca Signor, Jayesh A. Bafna, Rabia Sadir, Daphna Fenel, Elisabetta Boeri-Erba, Maria Bacia, Ninon Zala, Frédéric Laporte, Laurence Després, Martin Weik, Sébastien Boutet, Martin Rosenthal, Nicolas Coquelle, Manfred Burghammer, Duilio Cascio, Michael R. Sawaya, Mathias Winterhalter, Enrico Gratton, Irina Gutsche, Brian Federici, Jean-Luc Pellequer, Nicholas K. Sauter, Jacques-Philippe Colletier
Institutions: Centre National de la Recherche Scientifique, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, CEA Grenoble, Institut de Biologie Structurale, Lawrence Berkeley National Laboratory, SLAC National Accelerator Laboratory, Linac Coherent Light Source, European Synchrotron Radiation Facility, University of California, Riverside, Baylor University, Institute for Integrative Genome Biology, California Baptist University, Constructor University, Laboratoire d'Écologie Alpine, Université Grenoble Alpes, Institut Laue-Langevin, University of California, Los Angeles, University of California, Irvine
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,723
Funders: U.S. Department of Health and Human Services, Agence Nationale de la Recherche, European Synchrotron Radiation Facility, Centre National de la Recherche Scientifique, National Institutes of Health, Centers for Disease Control and Prevention, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, National Center for Research Resources
Awards: G113944070, CBH-EUR-GS, Time resolved insights into the photo-activation mechanism of OCP by combined ultrafast optical spectroscopy and serial femtosecond crystallography., G1825864493, Acquisition of a Scalable Storage Cluster for Data Intensive NIH Research, DIALS / CCTBX: Serial crystallography computational methods aimed at biomolecular function, G2300450775, Highly Improved Bacterial Larvicides for Vector Control, G336278089, In vivo crystallization: a novel strategy to obtain atomic-resolution insights into protein structure and dynamics at X-ray free-electron lasers and synchrotrons., G4532981089, G5011861974, G5570302943, G7184143679, G7403356940, PACIFIC SOUTHWEST REGIONAL CENTER OF EXCELLENCE FOR VECTOR-BORNE DISEASE RESEARCH AT THE UNIVERSITY OF CALIFORNIA, Infrastructure Française pour la Biologie Structurale Intégrée, A Synchrotron Radiation Structural Biology Resource, G9000266514
OpenAlex ID: W3009751908