Electron cryotomography of sub-cellular assemblies

Lead Research Organisation: University of Sheffield
Department Name: Molecular Biology and Biotechnology

Abstract

Future biotechnological applications of photosynthesis will require
bacteria that can harvest more of the solar spectrum than currently. That
way they could use more solar energy for making useful chemicals and
fuels. We are working on new pathways and combinations of pigments
in a photosynthetic bacterium that will exploit new parts of the solar
spectrum Ultimately we aim to convert a cell such as an E. coli into a
photosynthetic cell by engineering internal solar panels that allow them
to use sunlight for the first time. A crucial aspect of this work will involve
mapping in 3D molecular detail, arrangements of the protein pigments in
both native and engineered cells. To achieve this, we will exploit exciting
new developments in microscopy- cryo-electron tomography- that allow
us to visualise cells in unprecedented detail. We will create 3D images
of the photosynthetic machinery where we can even identify and locate
individual molecules. A crucial component of this project will involve the
development of new imaging methods that allow us to see the structures
more clearly. The project will involve combinations of biochemistry, use
and development of electron imaging approaches and computational

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011151/1 30/09/2015 29/09/2023
1801038 Studentship BB/M011151/1 30/09/2016 30/03/2021 Ainhao Dafis-Sagermendi
 
Description - Cryo-EM skills and expertise: sample preparation, independently using the microscopy facilities and using a range of programs for image processing
- Collaboration with group in Lisbon: greater understanding of bacterial endospore ultrastructure, especially in Clostridium difficile.
- Electron microscopy techniques to gain structural informations of the outermost layers of endospores. This is used to generate From 3D models of the surface layers.
- The outermost layer of Clostridium sporogenes endospores, a model organism for studying Clostridium botulinum, have been found to possess a highly resilient layer. This is similar to the outermost layers present in Bacillus.
Exploitation Route The studying of outermost layers in spores provides an insight into the host-pathogen interactions that take place with pathogens such as Clostridium difficile and Clostridium botulinum. This understanding can be used for the development of therapeutics or markers for treatment and detection pathogen respectively.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Assembly and composition of C.difficile endospore surface layers 
Organisation New University of Lisbon
Department António Xavier Institute of Chemical and Biological Technology
Country Portugal 
Sector Academic/University 
PI Contribution Electron microscopy expertise
Collaborator Contribution Cell samples and mutants.
Impact Manuscript in preparation
Start Year 2017