Time-Resolved Crystallography: Developing the Method and Solving the Mechanism of an Optogenetic Pump at Swissfel
- Doctoral Thesis
Rights / licenseIn Copyright - Non-Commercial Use Permitted
During my PhD studies at the ETH Zurich and the Paul Scherrer Institute I worked in the field of time-resolved serial femtosecond X-ray crystallography (TR-SFX), an emerging structural biology method that is performed at X-ray free-electron lasers (XFELs). The first advantage of XFELs is their provision of ultrafast 10-100 fs pulses, which allow radiation damage effects to be avoided, and obtaining structures from data collected at room temperature. The second advantage of XFELs comes upon installing a pump laser that induces protein conformational changes and allows obtaining crystallographic datasets at different times after protein activation. The protein that was subject to the study, Krokinobacter eikastus rhodopsin 2 (KR2), is the first discovered light-driven sodium pump. It was intriguing to elucidate the protein’s new sodium-pumping mechanism. It had been long debated if such a non-H+ cation pump could exist, because Na+ was supposed to be electrostatically repulsed from a positively charged retinal chromophore Schiff base. Due to the fact that outward pumping of Na+ silences neuronal signalling, KR2 is considered a next-generation tool for optogenetics. Show more
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ContributorsExaminer: Schertler, Gebhard F.X.
Examiner: Standfuss, Jörg
Examiner: Peter, Matthias
Examiner: Caflisch, Amedeo
SubjectXFELs, x-ray free electron lasers; Retinal; Membrane protein; MEMBRANE TRANSPORT, PERMEATION THROUGH MEMBRANES (BIOMEMBRANES); Membrane Transport; Protein crystallization; Protein crystallography; Lipidic cubic phase; Synchrotron; Serial crystallography; time-resolved serial femtosecond crystallography; SwissFEL; Sodium; Sodium transport; Pump-probe; Active transport; Bacteriorhodopsin; Bacterial proteins; Femtosecond X-ray pulses; Femtosecond; Protein Structure; Photoinduced energy transfer; Chromophore; Photocycle
Organisational unit03866 - Schertler, Gebhard F. / Schertler, Gebhard F.
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