Access Track 3:
Structure-function analysis of large protein complexes– from nano to micro
A central challenge in understanding the molecular mechanisms of health and disease is to learn how the structure of protein complexes and organelles gives rise to their function. The central underpinning of rational therapy design is that understanding the atomic and molecular structure of life’s machinery allows to predict their function and to interfere with it. However, it is clear that this process requires an integrated approach: high-resolution structure determination of isolated protein complexes must be combined with validation of the structure, and furthermore, the protein complexes’ interactions and, ultimately, their functions in a physiological context in situ. Only by directly combining in vitro and in situ imaging method, the structure can be determined in its functional context and the predicted function can be validated. Integrative modelling approaches can add to the understanding of protein complex functions in health, disease and after (drug) treatment. This access track offers valuable support in elucidating cellular mechanisms and in particular disease mechanisms and can thereby offer imminent impact for pharmaceutical applications and drug design. It is also most suitable for researchers working on imaging technology innovation and image data sharing through the development of new standards. Overall, all researchers across different scientific domains that have a need for access to integrated and correlative imaging technologies, data integration tools and repositories are invited to apply to Access Track 3.
Together, Euro-BioImaging, INSTRUCT, ISBE and ELIXIR have established an integrative access and service pipeline to methods from the nano- to the micrometer scale including atomic resolution three-dimensional (3D) information, 3D maps from electron and soft-xray microscopies, correlative light electron microscopy, super-resolution microscopy, functional imaging, data repositories, data standards, quantitative imaging analysis and data integration. If you are interested in getting access to some of these technologies - even if your scientific background is located far away from what we have described here! – do not hesitate, take your chance and submit your project proposal now!
In case of questions please contact the Access Track leader Frauke Leitner.
Example of an ongoing project:
Visualising picorna entry in the context of a novel viral entry pathway
"In our project, biosafety is a real issue. Without the help of CORBEL, it would have been much more difficult for us to get access to all the technologies we needed, because as an external scientist, you often just don’t know whom to talk to in a facility!"
Scientific interest: We have identified a lipid modifying enzyme factor that acts as host factor to facilitate the entry of picornavirus into human cells, using haploid genetic screening. This enzyme is not required for binding of the virus to the receptor, uptake into the cells or the formation of a membrane pore but instead, it acts in the understood next step; i.e. the efficient delivery of the genome into the cytoplasm. In the absence of this host factor a clearance mechanism was identified that was previously implicated in clearing bacterial pathogens.
Work plan: We now want to perform correlative light and electron microscopy (CLEM) at the Advanced Light Microscopy Facility, EMBL (Euro-BioImaging) to find the exact structure of the vesicles that release the viral genome into the cytoplasm. We also want to couple that with imaging cells by soft X-ray microscopy, which can specifically allow good visualisation of membranes in intact cells. The Instruct Image Processing Center (at the CNB-CSIC) (Instruct) will support us with the EM data acquisition planning and together with ELIXIR EMBL-EBI BioStudies database, they will provide software tools for data integration in an interlinked data repository.