Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade

Tetreau, Guillaume; Banneville, Anne-Sophie; Andreeva, Elena A.; Brewster, Aaron S.; Hunter, Mark S.; Sierra, Raymond G.; Teulon, Jean-Marie; Young, Iris D.; Burke, Niamh; Grünewald, Tilman A.; Beaudouin, Joël; Snigireva, Irina; Fernandez-Luna, Maria Teresa; Burt, Alister; Park, Hyun-Woo; Signor, Luca; Bafna, Jayesh A.; Sadir, Rabia; Fenel, Daphna; Boeri-Erba, Elisabetta; Bacia, Maria; Zala, Ninon; Laporte, Frédéric; Després, Laurence; Weik, Martin; Boutet, Sébastien; Rosenthal, Martin; Coquelle, Nicolas; Burghammer, Manfred; Cascio, Duilio; Sawaya, Michael R.; Winterhalter, Mathias; Gratton, Enrico; Gutsche, Irina; Federici, Brian; Pellequer, Jean-Luc; Sauter, Nicholas K.; Colletier, Jacques-Philippe

doi:10.1038/s41467-020-14894-w

Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade

Journal Article · Sun Mar 01 23:00:00 EST 2020 · Nature Communications

DOI:https://doi.org/10.1038/s41467-020-14894-w· OSTI ID:1617037

^[1]; Banneville, Anne-Sophie ^[1]; ^[1]; ^[2]; Hunter, Mark S. ^[3]; ^[2]; Teulon, Jean-Marie ^[1]; ^[2]; Burke, Niamh ^[1]; ^[4]; Beaudouin, Joël ^[1]; ^[4]; Fernandez-Luna, Maria Teresa ^[5]; ^[1]; Park, Hyun-Woo ^[6]; ^[1]; Bafna, Jayesh A. ^[7]; Sadir, Rabia ^[1]; Fenel, Daphna ^[1]; Boeri-Erba, Elisabetta ^[1] more »

Univ. Grenoble Alpes (France)
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
SLAC National Accelerator Lab., Menlo Park, CA (United States)
European Synchrotron Radiation Facility (ESRF), Grenoble (France)
Univ. of California, Riverside, CA (United States)
Univ. of California, Riverside, CA (United States); California Baptist Univ., Riverside, CA (United States)
Jacobs Univ., Bremen (Germany)
Institut Laue-Langevin, Grenoble (France)
Univ. of California, Los Angeles, CA (United States)
Univ. of California, Irvine, CA (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); Agence Nationale de la Recherche; U.S. Centers for Disease Control and Prevention; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1617037

Alternate ID(s):: OSTI ID: 1631638

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 11; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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