Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

Nogly, Przemyslaw; Panneels, Valérie; Nelson, Garrett; Gati, Cornelius; Kimura, Tetsunari; Milne, Christopher J.; Milathianaki, Despina; Kubo, Minoru; Wu, Wenting; Conrad, Chelsie E.; Coe, Jesse; Bean, Richard; Zhao, Yun; Båth, Petra; Dods, Robert; Harimoorthy, Rajiv; Beyerlein, Kenneth R.; Rheinberger, Jan; James, Daniel; DePonte, Daniel P.; Li, Chufeng; Sala, L.; Williams, Garth J.; Hunter, Mark S.; Koglin, Jason E.; Berntsen, Peter; Nango, Eriko; Iwata, So; Chapman, Henry N.; Fromme, Petra; Frank, Matthias; Boutet, Sébastien; Barty, Anton; White, Thomas A.; Weierstall, Uwe; Spence, John C. H.; Neutze, Richard; Schertler, Gebhard F. X.; Standfuss, Jörg

doi:10.1107/s2053273316099368

Cited by 5 publications

(8 citation statements)

References 49 publications

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“…This has meant that some details of the photochemical mechanism have remained poorly understood. A proof‐of‐principle study of bR dynamics was conducted at room temperature and resulted in a 2.3 Å resolution structure of an intermediate using lipidic‐cubic phase grown bR crystals [92]. A more extensive TR‐SFX study on bR showed that the photoisomerisation of retinal had already taken place prior to the earliest recorded time point at 16 ns (Fig.…”

Section: Natural Photoreceptorsmentioning

confidence: 99%

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A guide to time‐resolved structural analysis of light‐activated proteins

et al. 2021

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Dynamical changes in protein structures are essential for protein function and occur over femtoseconds to seconds timescales. X-ray free electron lasers have facilitated investigations of structural dynamics in proteins with unprecedented temporal and spatial resolution. Lightactivated proteins are attractive targets for time-resolved structural studies, as the reaction chemistry and associated protein structural changes can be triggered by short laser pulses. Proteins with different light-absorbing centres have evolved to detect light and harness photon energy to bring about downstream chemical and biological output responses. Following light-absorption, rapid chemical/small-scale structural changes are typically localised around the chromophore. These localised changes are followed by larger structural changes propagated throughout the photoreceptor/photocatalyst that enable the desired chemical and/or biological output response. Time-resolved serial-femtosecond crystallography (SFX) and solution scattering techniques enable direct visualisation of early chemical change in light-activated proteins on timescales previously inaccessible, whereas scattering gives access to slower timescales associated with more global structural change. Here, we review how advances in time-resolved SFX and solution scattering techniques have uncovered mechanisms of photochemistry and its coupling to output responses. We also provide a prospective on how these time-resolved structural approaches might impact on other photoreceptors/photoenzymes that have not yet been studied by these methods.

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Section: Natural Photoreceptorsmentioning

confidence: 99%

“…A resolution structure of an intermediate using lipidic-cubic phase grown bR crystals [92]. A more extensive TR-SFX study on bR showed that the photoisomerisation of retinal had already taken place prior to the earliest recorded time point at 16 ns (Fig.…”

Section: Rhodopsinsmentioning

confidence: 99%

A guide to time‐resolved structural analysis of light‐activated proteins

et al. 2021

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“…The secondary damage involves breaking of the disulfide bonds, decarboxylation of the acid group of amino acids, and other general effects such as an increase in the temperature factor, the increase in cell parameters, loss of high-resolution data, etc. [34]. When using pulses shorter than the Auger lifetime of atoms, the radiation damage can be reduced by outrunning the development of secondary electron-impact cascades [35,36].…”

Section: Free-electron Lasersmentioning

confidence: 99%

“…The continuous stream at high flow rate leads to a loss of precious samples between the X-ray pulses. To address this problem, high viscosity media such as lipids or mineral oil-based grease media are used [34][35][36]. A particular lipidic phase, the lipidic cubic phase (LCP) has previously been used to grow membrane protein crystals such as G-protein coupled receptors (GPCR) [37].…”

Section: Injector Systemsmentioning

confidence: 99%

Pump-Probe Time-Resolved Serial Femtosecond Crystallography at X-Ray Free Electron Lasers

2020

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With time-resolved crystallography (TRX), it is possible to follow the reaction dynamics in biological macromolecules by investigating the structure of transient states along the reaction coordinate. X-ray free electron lasers (XFELs) have enabled TRX experiments on previously uncharted femtosecond timescales. Here, we review the recent developments, opportunities, and challenges of pump-probe TRX at XFELs.

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“…Room temperature in situ data collection has also been implemented in various delivery methods and devices. For example, Nogly et al 54 used a lipidic cubic phase (LCP) injector in order to study the structure of the light-driven photon pump bacteriorhodopsin (bR) by serial femtosecond crystallography (SFX) using an XFEL source. The crystal structure of bR was solved to 2.3 Å resolution, demonstrating the compatibility of an LCP injector with time-resolved serial femtosecond crystallography (TR-SFX).…”

Section: Introductionmentioning

confidence: 99%