CoChat
Sign Up
PreprintCited by 9Insect Resistance and GeneticsInsect Pest Control StrategiesProtein Structure and Dynamics

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Structural journey of an insecticidal pore-forming protein targeting western corn rootworm

Open access
Marini, G., Poland, B., Leininger, C., Lukoyanova, N., Spielbauer, D., Barry, J., Altier, D., Lum, A., Scolaro, E., Pérez Ortega, C., Yalpani, N., Sandahl, G., Mabry, T., Klever, J., Nowatzki, T., Zhao, J-Z., Sethi, A., Kassa, A., Crane, V., Lu, A., Nelson, M.E., Eswar, N., Topf, M., Saibil, H.R.·2022·Nature Communications
DOI 10.1101/2022.10.12.511876PMC PMC10344926

Preprint — published as 10.1038/s41467-023-39891-7 · Paper page

Open this paper in CoChat — free accountOpen PDF

Abstract

Broad adoption of transgenic crops has revolutionized agriculture. However, resistance to insecticidal proteins by agricultural pests poses a continuous challenge to maintaining crop productivity and new proteins are urgently needed to replace existing transgenic traits. We identified an insecticidal membrane attack complex/perforin (MACPF) protein, Mpf2Ba1, with strong activity against western corn rootworm larvae and a novel site of action. By integrating X-ray crystallography, cryo-EM, and modelling, we determined monomeric, pre-pore and pore structures, revealing changes between structural states at atomic resolution. We discovered a monomer inhibition mechanism, a molecular “switch” associated with pre-pore activation/oligomerization upon gut fluid incubation and solved the highest resolution MACPF pore structure to-date. Our findings provide a mechanistic basis for Mpf2Ba1 effectiveness as an insecticidal protein with potential for biotechnology development. One-Sentence Summary: The molecular mechanism of an insecticidal protein is revealed through 3D structures of the three main pore formation states

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1101/2022.10.12.511876

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 78

  1. 10.1128/MMBR.00052-12 · DOI
  2. 10.1038/nrmicro.2015.3 · DOI
  3. 2019, Recent progress on the interaction between insects and Bacillus thuringiensis crops, Philosophical Transactions Royal Soc B, 374, 20180316, 10.1098/rstb.2018.0316 · DOI
  4. 2014, Key global environmental impacts of genetically modified (GM) crop use 1996–2012, Gm Crop Food, 5, 149, 10.4161/gmcr.28449 · DOI
  5. 2018, Farm income and production impacts of using GM crop technology 1996–2016, Gm Crop Food, 9, 1
  6. 10.1146/annurev.ento.54.110807.090434 · DOI
  7. N. Crickmore , C. Berry , S. Panneerselvam , R. Mishra , T. R. Connor , B. C. Bonning , A structure-based nomenclature for Bacillus thuringiensis and other bacteria-derived pesticidal proteins. J Invertebr Pathol, 107438 (2020). · DOI
  8. 2021, Resistance to Bt maize by Western Corn Rootworm: effects of pest biology, the pest–crop interaction and the agricultural landscape on resistance, Insects, 12, 136, 10.3390/insects12020136 · DOI
  9. 2016, Broad-spectrum resistance to Bacillus thuringiensis toxins by western corn rootworm (Diabrotica virgifera virgifera), Sci Rep-uk, 6, 27860, 10.1038/srep27860 · DOI
  10. 2010, Structure of a membrane-attack complex/perforin (MACPF) family protein from the human gut symbiont Bacteroides thetaiotaomicron, Acta Crystallogr Sect F Struct Biology Cryst Commun., 66, 1297, 10.1107/S1744309110023055 · DOI
  11. 2019, The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1, Nat Commun., 10, 4288, 10.1038/s41467-019-12279-2 · DOI
  12. 10.1126/sciadv.aax8286 · DOI
  13. 10.1038/353815a0 · DOI
  14. 1993, New folds for all-β proteins, Structure, 1, 217, 10.1016/0969-2126(93)90010-E · DOI
  15. 10.1126/science.1144706 · DOI
  16. 10.1073/pnas.1117597109 · DOI
  17. 2019, Structure–function characterization of an insecticidal protein GNIP1Aa, a member of an MACPF and β-tripod families, Proc National Acad Sci., 116, 201815547
  18. 10.1016/j.jip.2009.01.003 · DOI
  19. 2016, Giant MACPF/CDC pore forming toxins: A class of their own, Biochimica Et Biophysica Acta Bba-Biomembr., 1858, 475, 10.1016/j.bbamem.2015.11.017 · DOI
  20. 10.1007/s13238-016-0273-6 · DOI
  21. 10.1073/pnas.1423754112 · DOI
  22. 10.1371/journal.pbio.1002049 · DOI
  23. 10.1038/nsmb793 · DOI
  24. 10.1038/ncomms10588 · DOI
  25. 2019, An intermolecular π-stacking interaction drives conformational changes necessary to β-barrel formation in a pore-forming toxin, Mbio., 10, e01017
  26. 10.1016/0022-2836(94)90064-7 · DOI
  27. 10.1093/bioinformatics/bts152 · DOI
  28. 2002, The structure of bacterial outer membrane proteins, Biochimica Et Biophysica Acta Bba - Biomembr., 1565, 308, 10.1016/S0005-2736(02)00577-1 · DOI
  29. 10.1074/jbc.M112.430223 · DOI
  30. 10.1038/ncomms11293 · DOI
  31. 10.1126/science.aaf6056 · DOI
  32. 10.1111/pbi.12806 · DOI
  33. 10.1006/jmbi.1990.9999 · DOI · Paper page
  34. 2002, Digestibility of food allergens and nonallergenic proteins in simulated gastric fluid and simulated intestinal fluid—A Comparative Study, J Agr Food Chem., 50, 7154, 10.1021/jf020599h · DOI · Paper page
  35. 10.1007/s00299-014-1656-x · DOI
  36. 10.1603/0022-0493-98.1.1 · DOI
  37. 10.1038/90282 · DOI
  38. 10.1093/jee/tow049 · DOI
  39. 1984, Established Cell Lines from the Beetle, Diabrotica Undecimpunctata (Coleoptera: Chrysomelidae), Vitro., 20, 365, 10.1007/BF02619580 · DOI
  40. 2007, Identification of insect-selective and mammal-selective toxins from Parabuthus leiosoma venom, Toxicon, 50, 449, 10.1016/j.toxicon.2007.04.020 · DOI
  41. 2010, Features and development of Coot, Acta Crystallogr Sect D Biological Crystallogr., 66, 486, 10.1107/S0907444910007493 · DOI · Paper page
  42. 2011, REFMAC5 for the refinement of macromolecular crystal structures, Acta Crystallogr Sect D Biological Crystallogr., 67, 355, 10.1107/S0907444911001314 · DOI
  43. 10.1093/nar/gkw408 · DOI · Paper page
  44. 10.1002/pro.3235 · DOI · Paper page
  45. 10.1371/journal.pcbi.1004343 · DOI
  46. 10.1093/nar/28.1.235 · DOI · Paper page
  47. 2020, A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair, Nat Struct Mol Biol., 27, 71, 10.1038/s41594-019-0355-2 · DOI
  48. 1984, Cryo-electron microscopy of vitrified water, J Phys Colloq., C7, 85
  49. 10.1038/nmeth.4169 · DOI · Paper page
  50. 2021, DeepEMhancer: a deep learning solution for cryo-EM volume post-processing, Commun Biology, 4, 874, 10.1038/s42003-021-02399-1 · DOI
  51. 10.1016/j.jsb.2013.08.002 · DOI
  52. 10.1038/nmeth.4193 · DOI
  53. 10.1016/j.jsb.2015.08.008 · DOI
  54. 10.1016/j.jsb.2010.03.007 · DOI
  55. 10.1002/jcc.20084 · DOI
  56. 10.1016/j.jsb.2011.10.011 · DOI
  57. 10.1016/j.ymeth.2016.03.007 · DOI
  58. 2017, Recent developments in the CCP-EM software suite, Acta Crystallogr Sect D Struct Biology, 73, 469, 10.1107/S2059798317007859 · DOI
  59. 2018, ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps, Acta Crystallogr Sect D Struct Biology, 74, 519, 10.1107/S2059798318002425 · DOI
  60. 2018, New tools for the analysis and validation of cryo-EM maps and atomic models, Acta Crystallogr Sect D Struct Biology, 74, 814, 10.1107/S2059798318009324 · DOI
  61. 2019, Macromolecular structure determination using X‐rays, neutrons and electrons: recent developments in Phenix, Acta Crystallogr Sect D, 75, 861, 10.1107/S2059798319011471 · DOI · Paper page
  62. 2020, Procleave: Predicting protease-specific substrate cleavage sites by combining sequence and structural information, Genom Proteom Bioinform, 18, 52, 10.1016/j.gpb.2019.08.002 · DOI
  63. 10.1002/pro.3280 · DOI
  64. 10.1186/2046-1682-5-9 · DOI
  65. 10.1038/s41592-020-0731-1 · DOI
  66. 10.1107/S1600576715010092 · DOI
  67. 2010, MolProbity: all-atom structure validation for macromolecular crystallography, Acta Crystallogr Sect D Biological Crystallogr., 66, 12, 10.1107/S0907444909042073 · DOI · Paper page
  68. 10.1101/2021.12.16.472999 · DOI
  69. 2010, Estimating Diabrotica virgifera virgifera damage functions with field trial data: applying an unbalanced nested error component model, J Appl Entomol., 134, 409, 10.1111/j.1439-0418.2009.01487.x · DOI
  70. 2013, Validation of a nested error component model to estimate damage caused by corn rootworm larvae, J Appl Entomol., 137, 161, 10.1111/j.1439-0418.2012.01736.x · DOI
  71. 2011, Multiplexed protein quantification in maize leaves by liquid chromatography coupled with tandem mass spectrometry: an alternative tool to immunoassays for target protein analysis in genetically engineered crops, J Agr Food Chem., 59, 3551, 10.1021/jf104516r · DOI
  72. 10.1016/S0960-9822(00)80102-5 · DOI
  73. 10.1093/bioinformatics/btp033 · DOI
  74. 10.1126/sciadv.aaq0762 · DOI
  75. 10.1038/nature09518 · DOI
  76. 10.1074/jbc.M111.219766 · DOI
  77. 2018, The first transmembrane region of complement component-9 acts as a brake on its self-assembly, Nat Commun., 9, 3266, 10.1038/s41467-018-05717-0 · DOI
  78. 10.1074/jbc.M111.327809 · DOI

Metadata

Authors
Marini, G., Poland, B., Leininger, C., Lukoyanova, N., Spielbauer, D., Barry, J., Altier, D., Lum, A., Scolaro, E., Pérez Ortega, C., Yalpani, N., Sandahl, G., Mabry, T., Klever, J., Nowatzki, T., Zhao, J.-Z., Sethi, A., Kassa, A., Crane, V., Lu, A., Nelson, M. E., Eswar, N., Topf, M., & Saibil, H. R.
Affiliations
  • Leibniz Institute of Virology (LIV) (DE)
  • Corteva (United States) (US)
  • Syngenta (United States) (US)
  • Institute of Structural and Molecular Biology (GB)
Corresponding author
  • Nelson, M.E.
  • Eswar, N.
  • Topf, M.
  • Saibil, H.R.
ORCID
Venue
Nature Communications
Publisher
openRxiv
Published
2022-10-12
Volume
14
Issue
1
Pages
4171-4171
ISSN
2041-1723 (ISSN-L)
DOI
10.1101/2022.10.12.511876
PMC
PMC10344926
OpenAlex
W4384201122
Cited by
9
References
1
Citations by year
2026: 2 · 2025: 6 · 2023: 1
FWCI
0.80
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • BIROn (Birkbeck, University of London) (acceptedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • Desy Publications Database (Deutsches Elektronen-Synchrotron DESY) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • DESY Publication Database (PUBDB) (Deutsches Elektronen-Synchrotron)
Funders
  • U.S. Department of Agriculture
  • Iowa State University
  • Wellcome Trust (202679/Z/16/Z, Z/17/Z, 202679, 206166/Z/17/Z, 206166)
  • Corteva Agriscience
  • Agricultural Research Service
  • Plant Sciences Institute, Iowa State University
  • Leibniz-Gemeinschaft (W6/2018)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Insect Resistance and Genetics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Insect Pest Control Strategies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
western corn rootworm, insecticidal proteins, membrane attack complex, perforin, structural state, genetically modified crops, insect resistance, Coleoptera
MeSH terms
Animals, Coleoptera, Zea mays, Endotoxins, Insecticide Resistance, Insecticides, Larva, Pest Control, Biological, Animals, Genetically Modified, Plants, Genetically Modified, Perforin

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review