+Open data
-Basic information
Entry | Database: PDB / ID: 8b6w | |||||||||
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Title | Mpf2Ba1 pore | |||||||||
Components | Mpf2Ba1 | |||||||||
Keywords | TOXIN / Pore-forming protein / pore / oligomer / insecticidal / pesticide | |||||||||
Function / homology | MABP domain / MABP domain profile. / membrane-attack complex / perforin / Membrane attack complex/perforin (MACPF) domain profile. / MAC/Perforin domain / Membrane attack complex component/perforin (MACPF) domain / metal ion binding / cytoplasm / MACPF domain-containing protein Function and homology information | |||||||||
Biological species | Pseudomonas monteilii (bacteria) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.6 Å | |||||||||
Authors | Marini, G. / Poland, B. / Leininger, C. / Lukoyanova, N. / Spielbauer, D. / Barry, J. / Altier, D. / Lum, A. / Scolaro, E. / Perez Ortega, C. ...Marini, G. / Poland, B. / Leininger, C. / Lukoyanova, N. / Spielbauer, D. / Barry, J. / Altier, D. / Lum, A. / Scolaro, E. / Perez 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. | |||||||||
Funding support | United Kingdom, 2items
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Citation | Journal: Nat Commun / Year: 2023 Title: Structural journey of an insecticidal protein against western corn rootworm. Authors: Guendalina Marini / Brad Poland / Chris Leininger / Natalya Lukoyanova / Dan Spielbauer / Jennifer K Barry / Dan Altier / Amy Lum / Eric Scolaro / Claudia Pérez Ortega / Nasser Yalpani / ...Authors: Guendalina Marini / Brad Poland / Chris Leininger / Natalya Lukoyanova / Dan Spielbauer / Jennifer K Barry / Dan Altier / Amy Lum / Eric Scolaro / Claudia Pérez Ortega / Nasser Yalpani / Gary Sandahl / Tim Mabry / Jeffrey Klever / Timothy Nowatzki / Jian-Zhou Zhao / Amit Sethi / Adane Kassa / Virginia Crane / Albert L Lu / Mark E Nelson / Narayanan Eswar / Maya Topf / Helen R Saibil / Abstract: The broad adoption of transgenic crops has revolutionized agriculture. However, resistance to insecticidal proteins by agricultural pests poses a continuous challenge to maintaining crop productivity ...The 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 those utilized for existing transgenic traits. We identified an insecticidal membrane attack complex/perforin (MACPF) protein, Mpf2Ba1, with strong activity against the devastating coleopteran pest western corn rootworm (WCR) and a novel site of action. Using an integrative structural biology approach, we determined monomeric, pre-pore and pore structures, revealing changes between structural states at high resolution. We discovered an assembly inhibition mechanism, a molecular switch that activates pre-pore oligomerization upon gut fluid incubation and solved the highest resolution MACPF pore structure to-date. Our findings demonstrate not only the utility of Mpf2Ba1 in the development of biotechnology solutions for protecting maize from WCR to promote food security, but also uncover previously unknown mechanistic principles of bacterial MACPF assembly. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8b6w.cif.gz | 2.8 MB | Display | PDBx/mmCIF format |
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PDB format | pdb8b6w.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8b6w.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8b6w_validation.pdf.gz | 1.1 MB | Display | wwPDB validaton report |
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Full document | 8b6w_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 8b6w_validation.xml.gz | 247.8 KB | Display | |
Data in CIF | 8b6w_validation.cif.gz | 362.1 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/b6/8b6w ftp://data.pdbj.org/pub/pdb/validation_reports/b6/8b6w | HTTPS FTP |
-Related structure data
Related structure data | 15883MC 8b6uC 8b6vC C: citing same article (ref.) M: map data used to model this data |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 49961.504 Da / Num. of mol.: 21 / Source method: isolated from a natural source / Source: (natural) Pseudomonas monteilii (bacteria) / References: UniProt: A0A4Y8SM08 #2: Chemical | ChemComp-MG / Has ligand of interest | N | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Mpf2Ba1 21-mer pore / Type: COMPLEX / Entity ID: #1 / Source: NATURAL |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Pseudomonas monteilii (bacteria) |
Buffer solution | pH: 7.4 |
Specimen | Conc.: 0.6 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: TFS KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER |
Electron lens | Mode: OTHER / Nominal defocus max: 3300 nm / Nominal defocus min: 1500 nm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3D reconstruction | Resolution: 2.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 1413104 / Symmetry type: POINT |