Sampling Protein cOmplex Conformational Space with native top down Mass Spectrometry

Projektinformationen

SPOCkS MS

ID Finanzhilfevereinbarung: 759661

DOI 10.3030/759661

EK-Unterschriftsdatum 4 Oktober 2017

Startdatum 1 Januar 2018

Enddatum 30 September 2024

Finanziert unter

• EXCELLENT SCIENCE - European Research Council (ERC)

Gesamtkosten € 1 999 000,00

EU-Beitrag € 1 999 000,00

Koordiniert durch

LEIBNIZ-INSTITUT FUR VIROLOGIE
Germany

Ziel

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

Wissenschaftliches Gebiet

• natural sciences › computer and information sciences › software
• natural sciences › biological sciences › biochemistry › biomolecules › proteins
• natural sciences › chemical sciences › analytical chemistry › mass spectrometry
• natural sciences › biological sciences › molecular biology › structural biology
• natural sciences › physical sciences › optics › laser physics

Schlüsselbegriffe

• mass spectrometry
• fragmentation techniques
• labelling
• structural proteomics
• structural and biophysical virology
• free-electron lasers

Programm/Programme

• H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme

Thema/Themen

• ERC-2017-STG - ERC Starting Grant

Aufforderung zur Vorschlagseinreichung

ERC-2017-STG

Finanzierungsplan

ERC-STG - Starting Grant

Gastgebende Einrichtung

LEIBNIZ-INSTITUT FUR VIROLOGIE

• Netto-EU-Beitrag: € 1 999 000,00
• Adresse: MARTINISTRASSE 52 ; 20251 Hamburg ; Deutschland
• Region: Hamburg › Hamburg › Hamburg
• Aktivitätstyp: Research Organisations
• Gesamtkosten: € 1 999 000,00

Begünstigte (1)

LEIBNIZ-INSTITUT FUR VIROLOGIE

Deutschland

• Netto-EU-Beitrag: € 1 999 000,00
• Adresse: MARTINISTRASSE 52 ; 20251 Hamburg
• Region: Hamburg › Hamburg › Hamburg
• Aktivitätstyp: Research Organisations
• Gesamtkosten: € 1 999 000,00