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000626598 1001_ $$0P:(DE-H253)PIP1099525$$aKung, Jocky Chun Kui$$b0
000626598 245__ $$aX-ray Spectroscopy Meets Native Mass Spectrometry: Probing Gas-phase Protein Complexes
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000626598 520__ $$aGas-phase activation and dissociation studies of biomolecules, proteins and their non-covalent complexes using X-rays hold great promise for revealing new insights into the structure and function of biological samples. This is due to the unique properties of X-ray molecular interactions, such as site-specific and rapid ionization. In this perspective, we report and discuss the promise of first proof-of-principle studies of X-ray-induced dissociation of native biological samples ranging from small 17 kDa monomeric proteins up to large 808 kDa non-covalent protein assemblies conducted at a synchrotron (PETRA III) and a free-electron laser (FLASH2). A commercially available quadrupole time-of-flight mass spectrometer (Q-ToF2, Micromass/Waters), modified for high-mass analysis by MS Vision, was further adapted for integration with the open ports at the corresponding beamlines. The protein complexes were transferred natively into the gas phase via nano-electrospray ionization and subsequently probed by extreme ultraviolet (FLASH2) or soft X-ray (PETRA III) radiation, in either their folded state or following collision-induced activation in the gas phase. Depending on the size of the biomolecule and the activation method, protein fragmentation, dissociation, or enhanced ionization were observed. Additionally, an extension of the setup by ion mobility is described, which can serve as a powerful tool for structural separation of biomolecules prior to X-ray probing. The first experimental results are discussed in the broader context of current and upcoming X-ray sources, highlighting their potential for advancing structural biology in the future.
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000626598 7001_ $$0P:(DE-H253)PIP1018488$$aLu, Yinfei$$b18
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000626598 7001_ $$0P:(DE-H253)PIP1083974$$aSchamoni-Kast, Kira$$b21
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000626598 7001_ $$0P:(DE-HGF)0$$aSpringer, Sebastian$$b24
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000626598 7001_ $$0P:(DE-H253)PIP1018289$$aKierspel, Thomas$$b29$$eCorresponding author
000626598 7001_ $$0P:(DE-H253)PIP1014042$$aUetrecht, Charlotte$$b30$$eCorresponding author
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