Objective The main question to be addressed by SPOCk’S MS is how protein complex conformation adapts to local changes, such as processing of polyproteins, protein phosphorylation or conversion of substrates. While labelling strategies combined with mass spectrometry (MS), such as hydrogen deuterium exchange and hydroxyl footprinting, are very versatile in studying protein structure, these techniques are employed on bulk samples averaging over all species present. SPOCk’S MS will remedy these by studying the footprinting and therefore exposed surface area on conformation and mass selected species. Labelling still happens in solution avoiding gas phase associated artefacts. The labelling positions are then read out using newly developed top-down MS technology. Ultra-violet and free-electron lasers will be employed to fragment the protein complexes in the gas phase. In order to achieve the highest possible sequence and thus structural coverage, lasers will be complemented by additional dissociation and separation stages to allow MS^N. SPOCk’S MS will allow sampling conformational space of proteins and protein complexes and especially report about the transient nature of protein interfaces. Constraints derived in MS will be fed into a dedicated software pipeline to derive atomistic models. SPOCk’S MS will be used to study intracellular viral protein complexes, especially coronaviral replication/transcription complexes, which are highly flexible and often resist crystallisation and are barely accessible by conventional structural biology techniques.Objectives:- Integrate labelling with complex species selective native MS for time-resolved structural studies- Combine fragmentation techniques to maximise information content from MS- Develop software suite to analyse data and model protein complex structures based on MS constraints- Apply SPOCk’S MS to protein complexes of human pathogenic viruses Fields of science natural sciencescomputer and information sciencessoftwarenatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural scienceschemical sciencesanalytical chemistrymass spectrometrynatural sciencesbiological sciencesmolecular biologystructural biologynatural sciencesphysical sciencesopticslaser physics Keywords mass spectrometry fragmentation techniques labelling structural proteomics structural and biophysical virology free-electron lasers Programme(s) H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme Topic(s) ERC-2017-STG - ERC Starting Grant Call for proposal ERC-2017-STG See other projects for this call Funding Scheme ERC-STG - Starting Grant Coordinator LEIBNIZ-INSTITUT FUR VIROLOGIE Net EU contribution € 1 999 000,00 Address Martinistrasse 52 20251 Hamburg Germany See on map Region Hamburg Hamburg Hamburg Activity type Research Organisations Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 0,00 Beneficiaries (1) Sort alphabetically Sort by Net EU contribution Expand all Collapse all LEIBNIZ-INSTITUT FUR VIROLOGIE Germany Net EU contribution € 1 999 000,00 Address Martinistrasse 52 20251 Hamburg See on map Region Hamburg Hamburg Hamburg Activity type Research Organisations Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 0,00
