ArticleActa crystallographica. Section D, Biological crystallography2015
In meso in situ serial X-ray crystallography of soluble and membrane proteins.
Article in Acta crystallographica. Section D, Biological crystallography, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 63 papers.
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63 citing papers in PubMed, 111 citations in OpenAlex.
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- Improved detection of magnetic interactions in proteins based on long-lived coherences.Communications chemistry · 2024Article
- Review of serial femtosecond crystallography including the COVID-19 pandemic impact and future outlook.Structure (London, England : 1993) · 2023Review
- Micro-structured polymer fixed targets for serial crystallography at synchrotrons and XFELs.IUCrJ · 2023Article
- Native SAD phasing at room temperature.Acta crystallographica. Section D, Structural biology · 2022Article
- xia2.multiplex: a multi-crystal data-analysis pipeline.Acta crystallographica. Section D, Structural biology · 2022Article
- Acoustic levitation and rotation of thin films and their application for room temperature protein crystallography.Scientific reports · 2022Article
- Two states of a light-sensitive membrane protein captured at room temperature using thin-film sample mounts.Acta crystallographica. Section D, Structural biology · 2022Article
- Crystallographic Studies of Rhodopsins: Structure and Dynamics.Methods in molecular biology (Clifton, N.J.) · 2022Article
- An automated platform for structural analysis of membrane proteins through serial crystallography.Cell reports methods · 2021Article
- The crystal structure of bromide-boundeLife · 2021Article
- A fully automated crystallization apparatus for small protein quantities.Acta crystallographica. Section F, Structural biology communications · 2021Article
- Low-doseIUCrJ · 2020Article
- Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED.Structure (London, England : 1993) · 2020Article
- High-throughput in situ experimental phasing.Acta crystallographica. Section D, Structural biology · 2020Article
- 3D-printed holders forJournal of applied crystallography · 2020Article
- Recent advances in understanding prodrug transport through the SLC15 family of proton-coupled transporters.Biochemical Society transactions · 2020Review
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7 authors at 3 institutions in 3 countries.
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Abstract
The lipid cubic phase (LCP) continues to grow in popularity as a medium in which to generate crystals of membrane (and soluble) proteins for high-resolution X-ray crystallographic structure determination. To date, the PDB includes 227 records attributed to the LCP or in meso method. Among the listings are some of the highest profile membrane proteins, including the β2-adrenoreceptor-Gs protein complex that figured in the award of the 2012 Nobel Prize in Chemistry to Lefkowitz and Kobilka. The most successful in meso protocol to date uses glass sandwich crystallization plates. Despite their many advantages, glass plates are challenging to harvest crystals from. However, performing in situ X-ray diffraction measurements with these plates is not practical. Here, an alternative approach is described that provides many of the advantages of glass plates and is compatible with high-throughput in situ measurements. The novel in meso in situ serial crystallography (IMISX) method introduced here has been demonstrated with AlgE and PepT (alginate and peptide transporters, respectively) as model integral membrane proteins and with lysozyme as a test soluble protein. Structures were solved by molecular replacement and by experimental phasing using bromine SAD and native sulfur SAD methods to resolutions ranging from 1.8 to 2.8 Å using single-digit microgram quantities of protein. That sulfur SAD phasing worked is testament to the exceptional quality of the IMISX diffraction data. The IMISX method is compatible with readily available, inexpensive materials and equipment, is simple to implement and is compatible with high-throughput in situ serial data collection at macromolecular crystallography synchrotron beamlines worldwide. Because of its simplicity and effectiveness, the IMISX approach is likely to supplant existing in meso crystallization protocols. It should prove particularly attractive in the area of ligand screening for drug discovery and development.
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