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Mass Spectrometry for Single Particle Imaging of Dipole Oriented protein Complexes

Mass Spectrometry for Single Particle Imaging of Dipole Oriented protein Complexes

Objective

The European XFEL has just entered user operation. With its unparalleled peak brilliance and repetition rate, European XFEL has the potential to further applications in single particle imaging (SPI), thus far limited to large viral particles at X-ray Free-Electron Lasers (XFEL). SPI will allow imaging protein complexes without the need for crystallization. This eventually renders transient conformational states accessible for high resolution structural studies yielding molecular movies of biomolecular machines. A major bottleneck is the wealth of data required to reconstruct a single structure leading to long processing times. This is currently also a problem in electron microscopy (EM).
MS SPIDOC will overcome this data challenge by developing a native mass spectrometry (MS) system for sample delivery, named X-MS-I. It will provide mass and conformation selected biomolecules, which are oriented along their dipole axis upon imaging. This will enable structural reconstruction from much smaller datasets speeding up the analysis time tremendously. Moreover, the system features low sample consumption and a controlled low background easing pattern identification.
The main objectives of the project are:
• Deliver mass and conformation separated biomolecules for SPI.
• Orient proteins for SPI.
• Image protein complex unfolding
• Exploit potential of protein orientation for other applications
The MS SPIDOC consortium combines internationally leading expertise in different fields relevant to the project: Instrument design and development, computer simulations as well as working with biomolecules in the gas phase and on SPI are combined to implement the novel sample environment at the next generation XFEL facility. New components and methods will be opened to the market and thereby strengthen the European Research Area (ERA) and industry. This early stage high-risk project will give rise to a new technology with major impact on how to derive structures of biomolecules.
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Coordinator

HEINRICH-PETTE INSTITUT LEIBNIZ INSTITUT FUER EXPERIMENTELLE VIROLOGIE

Address

Martinistrasse 52
20251 Hamburg

Germany

Activity type

Research Organisations

EU Contribution

€ 499 125

Participants (7)

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EUROPEAN X-RAY FREE-ELECTRON LASERFACILITY GMBH

Germany

EU Contribution

€ 600 000

UNIVERSITAET GREIFSWALD

Germany

EU Contribution

€ 421 000

FASMATECH EPISTIMONIKI KAI TECHNOLOGIKI ANONYMI ETAIREIA

Greece

EU Contribution

€ 587 500

UNIVERSITE LYON 1 CLAUDE BERNARD

France

EU Contribution

€ 315 000

SPECTROMETRY VISION BV

Netherlands

EU Contribution

€ 432 500

UPPSALA UNIVERSITET

Sweden

EU Contribution

€ 512 500

THE UNIVERSITY OF MANCHESTER

United Kingdom

EU Contribution

€ 362 375

Project information

Grant agreement ID: 801406

Status

Ongoing project

  • Start date

    1 September 2018

  • End date

    31 August 2021

Funded under:

H2020-EU.1.2.1.

  • Overall budget:

    € 3 730 000

  • EU contribution

    € 3 730 000

Coordinated by:

HEINRICH-PETTE INSTITUT LEIBNIZ INSTITUT FUER EXPERIMENTELLE VIROLOGIE

Germany