Structure-factor analysis of femtosecond microdiffraction patterns from protein nanocrystals

Kirian, Richard A.; White, Thomas A.; Holton, James M.; Chapman, Henry N.; Fromme, Petra; Barty, Anton; Lomb, Lukas; Aquila, Andrew; Maia, Filipe R. N. C.; Martin, Andrew V.; Fromme, Raimund; Wang, Xiaoyu; Hunter, Mark S.; Schmidt, Kevin; Spence, John C. H.

doi:10.1107/s0108767310050981

Cited by 134 publications

(131 citation statements)

References 34 publications

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“…This improvement is more substantial when compared to other regions of integration. The optimal integration region can be affected by several factors, including crystal size and quality, pixel size and beam characteristics [36].…”

Section: Whole-pattern Fitting and Integration Analysismentioning

confidence: 99%

Analysis of Diffracted Intensities from Finite Protein Crystals with Incomplete Unit Cells

et al. 2017

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Developments in experimental techniques in micro electron diffraction and serial X-ray crystallography provide the opportunity to collect diffraction data from protein nanocrystals. Incomplete unit cells on the surfaces of protein crystals can affect the distribution of diffracted intensities for crystals with very high surface-to-volume ratios. The extraction of structure factors from diffraction data for such finite protein crystals sizes is considered here. A theoretical model for the continuous diffracted intensity distribution for data merged from finite crystals with two symmetry-related sub-units of the conventional unit cell is presented. This is used to extend a whole-pattern fitting technique to account for incomplete unit cells in the extraction of structure factor amplitudes. The accuracy of structure factor amplitudes found from this whole-pattern fitting technique and from an integration approach are evaluated.

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Section: Whole-pattern Fitting and Integration Analysismentioning

confidence: 99%

Analysis of Diffracted Intensities from Finite Protein Crystals with Incomplete Unit Cells

et al. 2017

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“…Following the rapid collection of several hundred thousand diffraction snapshots, Monte Carlo integration is applied to transform highly redundant partial reflections into structure factors [44]. The successful application of SFX was first demonstrated experimentally at low resolution, limited by the X-ray energy, on MPs crystallized in detergent solution [43] and in a liquid-like lipidic sponge phase [45].…”

Section: X-ray Free-electron Lasers and Femtosecond Crystallographymentioning

confidence: 99%

Femtosecond crystallography of membrane proteins in the lipidic cubic phase

Liu

Wacker

Wang

et al. 2014

Phil. Trans. R. Soc. B

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Despite recent technological advances in heterologous expression, stabilization and crystallization of membrane proteins (MPs), their structural studies remain difficult and require new transformative approaches. During the past two years, crystallization in lipidic cubic phase (LCP) has started gaining a widespread acceptance, owing to the spectacular success in highresolution structure determination of G protein-coupled receptors (GPCRs) and to the introduction of commercial instrumentation, tools and protocols. The recent appearance of X-ray free-electron lasers (XFELs) has enabled structure determination from substantially smaller crystals than previously possible with minimal effects of radiation damage, offering new exciting opportunities in structural biology. The unique properties of LCP material have been exploited to develop special protocols and devices that have established a new method of serial femtosecond crystallography of MPs in LCP (LCP-SFX). In this method, microcrystals are generated in LCP and streamed continuously inside the same media across the intersection with a pulsed XFEL beam at a flow rate that can be adjusted to minimize sample consumption. Pioneering studies that yielded the first room temperature GPCR structures, using a few hundred micrograms of purified protein, validate the LCP-SFX approach and make it attractive for structure determination of difficult-to-crystallize MPs and their complexes with interacting partners. Together with the potential of femtosecond data acquisition to interrogate unstable intermediate functional states of MPs, LCP-SFX holds promise to advance our understanding of this biomedically important class of proteins.

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“…Static SFX has been identified as the first 'killer app' of hard X-ray FELs [28,29]. How shall it be adapted to timeresolved experiments?…”

Section: Time-resolved Crystallography At Free-electron Lasersmentioning

confidence: 99%

Time-resolved crystallography and protein design: signalling photoreceptors and optogenetics

Moffat

2014

Phil. Trans. R. Soc. B

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Time-resolved X-ray crystallography and solution scattering have been successfully conducted on proteins on time-scales down to around 100 ps, set by the duration of the hard X-ray pulses emitted by synchrotron sources. The advent of hard X-ray free-electron lasers (FELs), which emit extremely intense, very brief, coherent X-ray pulses, opens the exciting possibility of time-resolved experiments with femtosecond time resolution on macromolecular structure, in both single crystals and solution. The X-ray pulses emitted by an FEL differ greatly in many properties from those emitted by a synchrotron, in ways that at first glance make time-resolved measurements of X-ray scattering with the required accuracy extremely challenging. This opens up several questions which I consider in this brief overview. Are there likely to be chemically and biologically interesting structural changes to be revealed on the femtosecond time-scale? How shall time-resolved experiments best be designed and conducted to exploit the properties of FELs and overcome challenges that they pose? To date, fast time-resolved reactions have been initiated by a brief laser pulse, which obviously requires that the system under study be light-sensitive. Although this is true for proteins of the visual system and for signalling photoreceptors, it is not naturally the case for most interesting biological systems. To generate more biological targets for time-resolved study, can this limitation be overcome by optogenetic, chemical or other means?

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Structure-factor analysis of femtosecond microdiffraction patterns from protein nanocrystals

Cited by 134 publications

References 34 publications

Analysis of Diffracted Intensities from Finite Protein Crystals with Incomplete Unit Cells

Analysis of Diffracted Intensities from Finite Protein Crystals with Incomplete Unit Cells

Femtosecond crystallography of membrane proteins in the lipidic cubic phase

Time-resolved crystallography and protein design: signalling photoreceptors and optogenetics

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