Skip to main content
U.S. Department of Energy
Office of Scientific and Technical Information

De novo determination of mosquitocidal Cry11Aa and Cry11Ba structures from naturally-occurring nanocrystals

Journal Article · · Nature Communications
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; more »; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; « less

Abstract

Cry11Aa and Cry11Ba are the two most potent toxins produced by mosquitocidal Bacillus thuringiensis subsp. israelensis and jegathesan , respectively. The toxins naturally crystallize within the host; however, the crystals are too small for structure determination at synchrotron sources. Therefore, we applied serial femtosecond crystallography at X-ray free electron lasers to in vivo-grown nanocrystals of these toxins. The structure of Cry11Aa was determined de novo using the single-wavelength anomalous dispersion method, which in turn enabled the determination of the Cry11Ba structure by molecular replacement. The two structures reveal a new pattern for in vivo crystallization of Cry toxins, whereby each of their three domains packs with a symmetrically identical domain, and a cleavable crystal packing motif is located within the protoxin rather than at the termini. The diversity of in vivo crystallization patterns suggests explanations for their varied levels of toxicity and rational approaches to improve these toxins for mosquito control.

Sponsoring Organization:
USDOE
Grant/Contract Number:
NONE; AC02-76SF00515
OSTI ID:
1878424
Alternate ID(s):
OSTI ID: 1887220
Journal Information:
Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United Kingdom
Language:
English

References (91)

The HMMER Web Server for Protein Sequence Similarity Search journal December 2017
Construction and Characterization of a Recombinant Bacillus thuringiensis subsp. israelensis Strain That Produces Cry11B journal July 2001
Bacillus thuringiensisCrylA(a) Insecticidal Toxin: Crystal Structure and Channel Formation journal December 1995
Improved production of the insecticidal CryIVD protein in Bacillus thuringiensis using cryIA(c) promoters to express the gene for an associated 20-kDa protein journal January 1995
Construction and characterization of the interdomain chimeras using Cry11Aa and Cry11Ba from Bacillus thuringiensis and identification of a possible novel toxic chimera journal September 2013
In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis journal May 1992
Statistics of local complexity in amino acid sequences and sequence databases journal June 1993
Rosetta3: An Object-Oriented Software Suite for the Simulation and Design of Macromolecules book January 2011
Structure of Cry2Aa Suggests an Unexpected Receptor Binding Epitope journal May 2001
CASS—CFEL-ASG software suite journal October 2012
Cry11Aa toxin from Bacillus thuringiensis binds its receptor in Aedes aegypti mosquito larvae through loop α-8 of domain II journal June 2005
Loop residues of the receptor binding domain of Bacillus thuringiensis Cry11Ba toxin are important for mosquitocidal activity journal May 2009
Current models of the mode of action of Bacillus thuringiensis insecticidal crystal proteins: A critical review journal September 2012
Contributions of 5′-UTR and 3′-UTR cis elements to Cyt1Aa synthesis in Bacillus thuringiensis subsp. israelensis journal October 2017
Crystal Structure of the Mosquito-larvicidal Toxin Cry4Ba and Its Biological Implications journal April 2005
Inference of Macromolecular Assemblies from Crystalline State journal September 2007
Cyt1Aa Toxin: Crystal Structure Reveals Implications for Its Membrane-Perforating Function journal November 2011
The adaptive gain integrating pixel detector AGIPD a detector for the European XFEL
  • Henrich, B.; Becker, J.; Dinapoli, R.
  • Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, Vol. 633 https://doi.org/10.1016/j.nima.2010.06.107
journal May 2011
Bayesian Deconvolution of Mass and Ion Mobility Spectra: From Binary Interactions to Polydisperse Ensembles journal April 2015
Multiple Receptors as Targets of Cry Toxins in Mosquitoes journal April 2011
Crystal structure of insecticidal δ-endotoxin from Bacillus thuringiensis at 2.5 Å resolution journal October 1991
Femtosecond X-ray protein nanocrystallography journal February 2011
De novo phasing with X-ray laser reveals mosquito larvicide BinAB structure journal September 2016
MMseqs2 enables sensitive protein sequence searching for the analysis of massive data sets journal October 2017
Automatic protein structure solution from weak X-ray data journal November 2013
Accurate macromolecular structures using minimal measurements from X-ray free-electron lasers journal March 2014
Concentric-flow electrokinetic injector enables serial crystallography of ribosome and photosystem II journal November 2015
Mass spectrometry of intact membrane protein complexes journal March 2013
Extreme amyloid polymorphism in Staphylococcus aureus virulent PSMα peptides journal August 2018
Megahertz data collection from protein microcrystals at an X-ray free-electron laser journal August 2018
Megahertz serial crystallography journal October 2018
Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade journal March 2020
Highly accurate protein structure prediction with AlphaFold journal July 2021
Crystallophore: a versatile lanthanide complex for protein crystallography combining nucleating effects, phasing properties, and luminescence journal January 2017
A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae journal January 2006
Biochemical characterization of Bacillus thuringiensis cytolytic toxins in association with a phospholipid bilayer journal February 1999
Bacillus thuringiensis subsp. israelensis Cyt1Aa synergizes Cry11Aa toxin by functioning as a membrane-bound receptor journal December 2005
Protein crystal structure obtained at 2.9 A resolution from injecting bacterial cells into an X-ray free-electron laser beam journal August 2014
Gas dynamic virtual nozzle for generation of microscopic droplet streams journal September 2008
CAST: an iterative algorithm for the complexity analysis of sequence tracts journal October 2000
STRAP: editor for STRuctural Alignments of Proteins journal April 2001
Fast and sensitive taxonomic assignment to metagenomic contigs journal March 2021
LCR-eXXXplorer: a web platform to search, visualize and share data for low complexity regions in protein sequences journal February 2015
The SWISS-PROT protein sequence data bank and its supplement TrEMBL in 1999 journal January 1999
Structural and functional characterization of DdrC, a novel DNA damage-induced nucleoid associated protein involved in DNA compaction journal July 2022
T-REX: a web server for inferring, validating and visualizing phylogenetic trees and networks journal June 2012
Alignment-Annotator web server: rendering and annotating sequence alignments journal May 2014
HMMER web server: 2018 update journal June 2018
The EMBL-EBI search and sequence analysis tools APIs in 2019 journal April 2019
Low complexity regions in the proteins of prokaryotes perform important functional roles and are highly conserved journal September 2019
Phaser crystallographic software journal July 2007
Indexing in single-crystal diffractometry with an obstinate list of reflections journal April 1992
MOLREP an Automated Program for Molecular Replacement journal December 1997
Structure of the insecticidal bacterial δ-endotoxin Cry3Bb1 of Bacillus thuringiensis journal July 2001
Refinement of severely incomplete structures with maximum likelihood in BUSTER–TNT journal November 2004
Coot model-building tools for molecular graphics journal November 2004
Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions journal November 2004
The integration of macromolecular diffraction data journal December 2005
Iterative model building, structure refinement and density modification with the PHENIX AutoBuild wizard journal December 2007
ALINE : a WYSIWYG protein-sequence alignment editor for publication-quality alignments journal April 2009
Overview of the CCP 4 suite and current developments journal March 2011
REFMAC 5 for the refinement of macromolecular crystal structures journal March 2011
Completion of autobuilt protein models using a database of protein fragments journal March 2012
Towards automated crystallographic structure refinement with phenix.refine journal March 2012
New Python-based methods for data processing journal June 2013
Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams journal April 2015
Protein crystal structure determination with the crystallophore, a nucleating and phasing agent journal June 2019
The Coherent X-ray Imaging instrument at the Linac Coherent Light Source journal April 2015
Serial femtosecond crystallography: the first five years journal February 2015
XGANDALF – extended gradient descent algorithm for lattice finding journal August 2019
Diffraction-geometry refinement in the DIALS framework journal March 2016
TakeTwo: an indexing algorithm suited to still images with known crystal parameters journal July 2016
Distributed computing for macromolecular crystallography journal February 2018
DIALS : implementation and evaluation of a new integration package journal February 2018
Processing serial crystallography data with CrystFEL : a step-by-step guide journal January 2019
Bacillus thuringiensis ssp. israelensis Cyt1Aa enhances activity of Cry11Aa toxin by facilitating the formation of a pre-pore oligomeric structure: Cyt1Aa facilitates oligomerization of Cry11Aa toxin journal August 2007
Bacillus thuringiensis insecticidal three-domain Cry toxins: mode of action, insect resistance and consequences for crop protection journal January 2013
Atomic structures of low-complexity protein segments reveal kinked β sheets that assemble networks journal February 2018
Accurate prediction of protein structures and interactions using a three-track neural network journal July 2021
Binding of Cyt1Aa and Cry11Aa Toxins of Bacillus thuringiensis Serovar israelensis to Brush Border Membrane Vesicles of Tipula paludosa (Diptera: Nematocera) and Subsequent Pore Formation journal June 2007
Oligomerization of Cry11Aa from Bacillus thuringiensis Has an Important Role in Toxicity against Aedes aegypti journal December 2009
Structure of the Functional Form of the Mosquito Larvicidal Cry4Aa Toxin from Bacillus thuringiensis at a 2.8-Angstrom Resolution journal May 2006
Cloning and expression of a novel toxin gene from Bacillus thuringiensis subsp. jegathesan encoding a highly mosquitocidal protein journal December 1995
DeStripe: frequency-based algorithm for removing stripe noises from AFM images journal January 2011
Aromatic clusters in protein–protein and protein–drug complexes journal May 2020
Functional Analysis of Two Processed Fragments of Bacillus thuringiensis Cry11A Toxin journal January 2004
Dominant Negative Phenotype of Bacillus thuringiensis Cry1Ab, Cry11Aa and Cry4Ba Mutants Suggest Hetero-Oligomer Formation among Different Cry Toxins journal May 2011
A theoretical model of the tridimensional structure of Bacillus thuringiensis subsp. medellin Cry 11Bb toxin deduced by homology modelling journal April 2001
Comparing Median Lethal Concentration Values Using Confidence Interval Overlap or Ratio Tests journal January 2006
Bacillus Thuringiensis Serovariety Israelensis and Bacillus Sphaericus for Mosquito Control journal July 2007
Structural Insights into Bacillus thuringiensis Cry, Cyt and Parasporin Toxins journal September 2014

Similar Records

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 · OSTI ID:1617037

Related Subjects