Molecular analysis of the interaction of Bin pesticidal protein with its receptor
Lead Research Organisation:
Cardiff University
Department Name: School of Biosciences
Abstract
Bacillus and Lysinibacillus bacteria are able to produce a range of insecticidal toxins that are highly specific for narrow ranges of target insects. This makes them attractive agents for use as biopesticides for the control of pest insects. Specificity is determined by recognition of receptors on target cells but the molecular details of toxin-receptor interactions have not been elucidated for any of these toxins. The Bin toxin of Lysinibacillus sphaericus is selectively active against Culex and Anopheles mosquitoes and the receptor protein, an alpha glycosidase, has been identified. Recognition of this receptor is mediated by the BinB protein and a structure for the latter has been elucidated.
The project will investigate the interaction of BinB and the Cqm1 receptor from Culex quinquefasciatus. The receptor will be produced in recombinant form for crystallization trials to solve its structure. Complexes of BinB and Cqm1 will be produced for further crystal trials. Regions of the proteins interacting will also be probed by modelling. Cluspro and Rosetta will be used to predict interacting regions. Mutation of toxin and receptor will be used to probe the importance of these regions. The incorporation of unnatural amino acids amenable to photoactivatable cross linking will allow the trapping of complexes. The effects of mutations will be tested in cells in culture and in toxicity bioassays monitoring killing of mosquito larvae.
The student appointed will learn techniques of molecular biology, protein biochemistry, cell culture and structural analysis.
The project will investigate the interaction of BinB and the Cqm1 receptor from Culex quinquefasciatus. The receptor will be produced in recombinant form for crystallization trials to solve its structure. Complexes of BinB and Cqm1 will be produced for further crystal trials. Regions of the proteins interacting will also be probed by modelling. Cluspro and Rosetta will be used to predict interacting regions. Mutation of toxin and receptor will be used to probe the importance of these regions. The incorporation of unnatural amino acids amenable to photoactivatable cross linking will allow the trapping of complexes. The effects of mutations will be tested in cells in culture and in toxicity bioassays monitoring killing of mosquito larvae.
The student appointed will learn techniques of molecular biology, protein biochemistry, cell culture and structural analysis.
People |
ORCID iD |
| Colin Berry (Primary Supervisor) | |
| Lainey Williamson (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M009122/1 | 01/10/2015 | 31/03/2024 | |||
| 2194445 | Studentship | BB/M009122/1 | 01/10/2019 | 30/09/2023 | Lainey Williamson |
| Description | Several protein structures of insecticidal proteins have been elucidated using serial femtosecond crystallography. The interaction of the binding component of the Bin pesticidal protein, BinB, with its receptor protein, Cqm1, has been modelled using computational techniques. Increased research capability of the PhD student involved in the award has been generated from extensive training in the field of insecticidal proteins and protein crystallography. |
| Exploitation Route | The agrotech industry is anticipated to benefit from an increased understanding of the structure of insecticidal proteins, which is expected to underpin protein engineering studies aimed at generating enhanced forms for use as bioinsecticides and in transgenic crops. This work has the potential to lead to toxin commercialisation and patentable products, which in turn will benefit the general public in the form of better food security and an improved management of disease transmitting insects, such as mosquitoes. This project has provided the graduate researcher with a broad range of transferable research skills at the biological, chemical, and physical cross-disciplinary interface. This will greatly support their career progression into academic or industrial research roles. |
| Sectors | Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology |
| Title | Tpp49Aa1 crystal structure - pH 11 |
| Description | Crystal protein structure solved using serial femtosecond crystallography |
| Type Of Material | Database/Collection of data |
| Year Produced | 2023 |
| Provided To Others? | No |
| Impact | Novel protein structure |
| Title | Tpp49Aa1 crystal structure - pH 3 |
| Description | Crystal protein structure solved using serial femtosecond crystallography - PDB 8BEX |
| Type Of Material | Database/Collection of data |
| Year Produced | 2023 |
| Provided To Others? | No |
| Impact | Novel protein structure |
| Title | Tpp49Aa1 crystal structure - pH 7 (1.62 Å) |
| Description | Crystal protein structure solved using serial femtosecond crystallography - PDB 8BEY |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | No |
| Impact | Novel protein structure |
| Title | Tpp49Aa1 crystal structure - pH 7 (2.2 Å) |
| Description | Crystal protein structure solved using serial femtosecond crystallography - PDB 7QA1 |
| Type Of Material | Database/Collection of data |
| Year Produced | 2021 |
| Provided To Others? | No |
| Impact | Novel protein structure |
| URL | https://www.rcsb.org/structure/unreleased/7QA1 |
| Title | Tpp80Aa1 structure |
| Description | PDB submission dataset 8BAD |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| Impact | Publication |
| URL | https://www.rcsb.org/structure/8BAD |
| Description | CalBap |
| Organisation | California Baptist University |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Provision of toxin preparations for bioassays |
| Collaborator Contribution | Bioassays carried out and analysed. |
| Impact | Input to our PNAS manuscript submission |
| Start Year | 2022 |
| Description | EuXFEL and DESY |
| Organisation | Deutsches Electronen-Synchrotron (DESY) |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | Sample preparation for serial femtosecond crystallography studies, diffraction data analysis and structure solution of several protein structures |
| Collaborator Contribution | Diffraction data collection, processing, and analysis |
| Impact | - Award of three beamtime allocations at the European XFEL - Full datasets for seven novel protein structures (several manuscripts in preparation) - International conferences presentations: Williamson, L.J. 2023: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Invited speaker, Structural Biology Seminar Series. Walter and Eliza Hall Institute of Medical Research. Melbourne, Australia. Williamson, L.J. 2023: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Oral presentation. Lorne Conference on Protein Structure and Function. Lorne, Australia. Williamson, L.J. 2022: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Poster presentation. International Congress on Invertebrate Pathology and Microbial Control & Annual Meeting of the Society for Invertebrate Pathology. Online. Williamson, L.J. 2022: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Poster presentation. South West Structural Biology Consortium. Bristol, UK. Williamson, L.J. 2021: "Crystal structure of Lysinibacillus sphaericus Tpp49 solved using serial femtosecond crystallography". Poster presentation. International Congress on Invertebrate Pathology and Microbial Control & Annual Meeting of the Society for Invertebrate Pathology. Online. |
| Start Year | 2019 |
| Description | EuXFEL and DESY |
| Organisation | European XFEL |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | Sample preparation for serial femtosecond crystallography studies, diffraction data analysis and structure solution of several protein structures |
| Collaborator Contribution | Diffraction data collection, processing, and analysis |
| Impact | - Award of three beamtime allocations at the European XFEL - Full datasets for seven novel protein structures (several manuscripts in preparation) - International conferences presentations: Williamson, L.J. 2023: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Invited speaker, Structural Biology Seminar Series. Walter and Eliza Hall Institute of Medical Research. Melbourne, Australia. Williamson, L.J. 2023: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Oral presentation. Lorne Conference on Protein Structure and Function. Lorne, Australia. Williamson, L.J. 2022: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Poster presentation. International Congress on Invertebrate Pathology and Microbial Control & Annual Meeting of the Society for Invertebrate Pathology. Online. Williamson, L.J. 2022: "Using serial femtosecond crystallography to solve the structures of pesticidal proteins from native crystals". Poster presentation. South West Structural Biology Consortium. Bristol, UK. Williamson, L.J. 2021: "Crystal structure of Lysinibacillus sphaericus Tpp49 solved using serial femtosecond crystallography". Poster presentation. International Congress on Invertebrate Pathology and Microbial Control & Annual Meeting of the Society for Invertebrate Pathology. Online. |
| Start Year | 2019 |
| Description | Federici |
| Organisation | University of California, Riverside |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Provision of toxin preparations for bioassays |
| Collaborator Contribution | Bioassays carried out and analysed. |
| Impact | Input to our PNAS manuscript submission |
| Start Year | 2022 |
| Description | Monash University |
| Organisation | Monash University |
| Country | Australia |
| Sector | Academic/University |
| PI Contribution | Protein sample preparation and characterisation of pore formation |
| Collaborator Contribution | CryoEM studies |
| Impact | - Award of BBSRC partnering award leading to research exchange between laboratories |
| Start Year | 2019 |
| Description | Interview for press release (Deutsches Elektronen-Synchrotron DESY) |
| Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Interview for press release by the Deutsches Elektronen-Synchrotron DESY (Hamburg, Germany) - associated with using serial femtosecond crystallography to solve the structure of Tpp49Aa1 from natural crystals |
| Year(s) Of Engagement Activity | 2023 |
| Description | Invited speaker at Walter and Eliza Hall Institute of Medical Research Institute |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Presentation on 'Using serial femtosecond crystallography to solve the structures of pesticidal proteins from their native crystals' (Walter and Eliza Hall Institute of Medical Research Institute - Melbourne, Australia) |
| Year(s) Of Engagement Activity | 2023 |
| Description | Presentation to scientific advisory board |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Industry/Business |
| Results and Impact | Presentation on 'using computational modelling techniques to improve the agronomic performance of target genes' to scientific advisory board meeting hosted by Agritech industry company |
| Year(s) Of Engagement Activity | 2023 |
| Description | Q&A session for sixth form school students |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Q&A session aimed at encouraging sixth form school students to pursue careers in STEM subjects |
| Year(s) Of Engagement Activity | 2022 |
| Description | University open days |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Schools |
| Results and Impact | School of Biosciences university open day for sixth-form students |
| Year(s) Of Engagement Activity | 2020,2021,2022 |