Quantitative reactivity profiling of the cytochrome P450 superfamily
Lead Research Organisation:
University of Oxford
Department Name: Interdisciplinary Bioscience DTP
Abstract
"Cytochromes P450 (P450s) form a large and particularly diverse superfamily of enzymes in plants. P450s play an important role in the biosynthesis of many structural components, plant hormones and defence compounds and are key enzymes in the detoxification of agrochemicals 1. Many efforts have been taken in order to identify the specificity of these enzymes and understand their biological function. However, just a minor number of P450 substrates have been identified and thus, there still remain many P450s to be functionally characterised 2. In this research we aim to use a distinct and more global approach such is activity-based protein profiling (ABPP) to monitor, identify and characterise the activity of P450 enzymes within complex plant proteomes. The aim of this project is to establish a quantitative reactivity profiling of the cytochrome P450 superfamily in plants and apply this technology to study the role of P450s in specific plant phenomena. In short, firstly we will exploit used and novel P450 probes to generate a new suite of activity-based probes able to quantitatively profile P450s in the model plant Arabidiopsis thaliana. Subsequently, we will implement this distinct tool to identify and monitor the activity of P450s in A. thaliana-Pseudomonas syringae interaction and different agrochemical treatments.
BBSRC priority areas addressed:
- Agriculture and Food Security.
- Food, nutrition and health.
- New strategic approaches to industrial biotechnology.
- Sustainably enhancing agricultural production.
- Global food security."
BBSRC priority areas addressed:
- Agriculture and Food Security.
- Food, nutrition and health.
- New strategic approaches to industrial biotechnology.
- Sustainably enhancing agricultural production.
- Global food security."
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011224/1 | 01/10/2015 | 31/03/2024 | |||
| 1876717 | Studentship | BB/M011224/1 | 01/01/2017 | 01/07/2021 |
| Description | Activity-based protein profiling, a chemical-proteomic technique, has been introduced in the field of herbicide research by the establishment and validation of glutathione S-transferase profiling (herbicide detoxification enzymes) in plants. Furthermore, activity-based protein profiling of cytochrome P450s has been successfully implemented in the fungi Zymoseptoria tritici, making P450 profiling a powerful technique to study fungicide detoxification. In plants, P450 probes can just label transiently overexpressed P450s, suggesting that probes are suitable for plant studies but the technique needs further optimization and development. |
| Exploitation Route | With the GST-G probe glutathione S-transferases can be labelled and this chemical probe can be used to study the role of GSTs in different biotic or abiotic plant stresses. The P450-targeting probes can be used to study the role of P450s in fungicide detoxification. |
| Sectors | Agriculture, Food and Drink,Environment |
| Description | Anne Clements Travel Grant , Somerville College |
| Amount | £340 (GBP) |
| Organisation | University of Oxford |
| Department | Somerville College |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 07/2018 |
| End | 07/2018 |
| Description | Activity-based protein profiling of serine hydrolases in carnivorous plants |
| Organisation | University of Oxford |
| Department | Oxford Botanic Garden |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We perform serine hydrolase protein profiling to study serine hydrolase activities in carnivorous plants deliberately stimulated with insect prey |
| Collaborator Contribution | Oxford Botanic Garden allow us to access the carnivorous plant collection in order to perform the experiments |
| Impact | no impact yet |
| Start Year | 2018 |
| Description | Expanding cytochrome P450s and glutathione S-transferases profiling in plants by synthesising novel activity-based probes |
| Organisation | University of Oxford |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | In this partnership we tested the chemical probes synthesized by our collaborators in plant tissues. |
| Collaborator Contribution | The collaborators synthesized new chemical probes in order to detect the activity of two important family of proteins involved in herbicide detoxification (P450s and GSTs). |
| Impact | No impact yet |
| Start Year | 2018 |
| Description | Co-organization of the Target Discovery Using Chemical Probes symposium |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | We hosted the 11th International Conference on Activity-based Protein Profiling (Target Discovery Using Chemical Probes symposium) in Oxford: 104 participants, 30 presentations and 35 posters. |
| Year(s) Of Engagement Activity | 2018 |
| Description | SuperScience day at the Natural Museum of Oxford |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | We design an activity for the Natural Museum Science Day where we wanted to make children aware about how plants interact with other small organisms (bacteria and fungi) and that some of these tiny organisms can help the plants to grow but also can make them feel sick. |
| Year(s) Of Engagement Activity | 2018 |