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Updated: May 9, 2025
07:33
Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
Published on: October 15, 2018
14.1K
X-ray spectroscopy meets native mass spectrometry: probing gas-phase protein complexes
Jocky C K Kung1,2,3, Alan Kádek1,4,5, Knut Kölbel6
1CSSB Centre for Structural Systems Biology, Deutsches Elektronen-Synchrotron DESY & Leibniz Institute of Virology (LIV) & University of Lübeck, Notkestraße 85, 22607 Hamburg, Germany. thomas.kierspel@desy.de.
Physical Chemistry Chemical Physics : PCCP
|April 30, 2025
Summary
X-ray-induced dissociation of proteins and their complexes in the gas phase offers new insights into biological structure and function. This study demonstrates proof-of-principle experiments using synchrotron and free-electron laser sources.
Area of Science:
- Structural biology
- Biophysics
- Mass spectrometry
Background:
- X-ray interactions with biomolecules provide unique insights into structure and function.
- Site-specific and rapid ionization are key properties of X-ray molecular interactions.
- Gas-phase studies of proteins and their complexes are emerging as powerful tools.
Purpose of the Study:
- To report and discuss proof-of-principle studies of X-ray-induced dissociation of native biological samples.
- To investigate X-ray-induced fragmentation, dissociation, and ionization of proteins and protein assemblies.
- To highlight the potential of X-ray sources for advancing structural biology.
Main Methods:
- Native protein complexes were transferred into the gas phase via nano-electrospray ionization.
- Samples were probed using extreme ultraviolet (FLASH2) or soft X-ray (PETRA III) radiation.
- A modified quadrupole time-of-flight mass spectrometer was adapted for synchrotron and free-electron laser beamlines.
Main Results:
- X-ray-induced dissociation was observed for biomolecules ranging from 17 kDa proteins to 808 kDa non-covalent assemblies.
- Protein fragmentation, dissociation, and enhanced ionization were dependent on biomolecule size and activation method.
- Ion mobility was integrated for structural separation prior to X-ray probing.
Conclusions:
- X-ray-induced dissociation is a promising technique for studying native biological samples in the gas phase.
- The study demonstrates the feasibility of using synchrotron and free-electron laser sources for these investigations.
- Future X-ray sources hold significant potential for advancing structural biology research.
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