ArticleProceedings of the National Academy of Sciences of the United States of America2016
Membrane protein structure determination by SAD, SIR, or SIRAS phasing in serial femtosecond crystallography using an iododetergent.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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28 citing papers in PubMed, 49 citations in OpenAlex.
- A simple and practical approach for protein serial crystallography using grease matrix and large-area support film.Journal of synchrotron radiation · 2026Article
- Advances in Crystallography and Cryo-Electron Microscopy for the Study of Membrane Proteins.Advances in experimental medicine and biology · 2026Review
- Tracking Protein Motions using Serial Femtosecond Crystallography with X-Ray Free-Electron Laser.Current protocols · 2025Article
- A versatile approach to high-density microcrystals in lipidic cubic phase for room-temperature serial crystallography.Journal of applied crystallography · 2023Article
- Single-support serial isomorphous replacement phasing.Acta crystallographica. Section D, Structural biology · 2022Article
- Ultrafast isolated molecule imaging without crystallization.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Crystallization of Microbial Rhodopsins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Crystal structure of CmABCB1 multi-drug exporter in lipidic mesophase revealed by LCP-SFX.IUCrJ · 2022Article
- Article
- Heavy Atom Detergent/Lipid Combined X-ray Crystallography for Elucidating the Structure-Function Relationships of Membrane Proteins.Membranes · 2021Review
- Discerning best practices in XFEL-based biological crystallography - standards for nonstandard experiments.IUCrJ · 2021Review
- Dynamic Structural Biology Experiments at XFEL or Synchrotron Sources.Methods in molecular biology (Clifton, N.J.) · 2021Review
- Article
- Experimental phasing with vanadium and application to nucleotide-binding membrane proteins.IUCrJ · 2020Article
- Viscosity-adjustable grease matrices for serial nanocrystallography.Scientific reports · 2020Article
- High-viscosity sample-injection device for serial femtosecond crystallography at atmospheric pressure.Journal of applied crystallography · 2019Article
- SAD phasing of XFEL data depends critically on the error model.Acta crystallographica. Section D, Structural biology · 2019Article
- An outlook on using serial femtosecond crystallography in drug discovery.Expert opinion on drug discovery · 2019Review
- Multiscale Simulations of Biological Membranes: The Challenge To Understand Biological Phenomena in a Living Substance.Chemical reviews · 2019Article
- Serial femtosecond crystallography at the SACLA: breakthrough to dynamic structural biology.Biophysical reviews · 2018Review
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31 authors at 7 institutions in 2 countries.
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No grant is acknowledged in the PubMed record.
Abstract
The 3D structure determination of biological macromolecules by X-ray crystallography suffers from a phase problem: to perform Fourier transformation to calculate real space density maps, both intensities and phases of structure factors are necessary; however, measured diffraction patterns give only intensities. Although serial femtosecond crystallography (SFX) using X-ray free electron lasers (XFELs) has been steadily developed since 2009, experimental phasing still remains challenging. Here, using 7.0-keV (1.771 Å) X-ray pulses from the SPring-8 Angstrom Compact Free Electron Laser (SACLA), iodine single-wavelength anomalous diffraction (SAD), single isomorphous replacement (SIR), and single isomorphous replacement with anomalous scattering (SIRAS) phasing were performed in an SFX regime for a model membrane protein bacteriorhodopsin (bR). The crystals grown in bicelles were derivatized with an iodine-labeled detergent heavy-atom additive 13a (HAD13a), which contains the magic triangle, I3C head group with three iodine atoms. The alkyl tail was essential for binding of the detergent to the surface of bR. Strong anomalous and isomorphous difference signals from HAD13a enabled successful phasing using reflections up to 2.1-Å resolution from only 3,000 and 4,000 indexed images from native and derivative crystals, respectively. When more images were merged, structure solution was possible with data truncated at 3.3-Å resolution, which is the lowest resolution among the reported cases of SFX phasing. Moreover, preliminary SFX experiment showed that HAD13a successfully derivatized the G protein-coupled A2a adenosine receptor crystallized in lipidic cubic phases. These results pave the way for de novo structure determination of membrane proteins, which often diffract poorly, even with the brightest XFEL beams.
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