ArticleActa crystallographica. Section D, Structural biology2016
A pipeline for structure determination of in vivo-grown crystals using in cellulo diffraction.
Article in Acta crystallographica. Section D, Structural biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 42 citations in OpenAlex.
- Review
- Single-cell structural biology with intracellular electron crystallography.Nature communications · 2026Article
- Intracellular protein crystallization in living insect cells.FEBS open bio · 2025Article
- High-throughput structure determination of an intrinsically disordered protein using cell-free protein crystallization.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- A streamlined approach to structure elucidation using in cellulo crystallized recombinant proteins, InCellCryst.Nature communications · 2024Article
- Atomic structure of a nudivirus occlusion body protein determined from a 70-year-old crystal sample.Nature communications · 2023Article
- Cell-free protein crystallization for nanocrystal structure determination.Scientific reports · 2022Article
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- Can (We Make)Toxins · 2021Review
- Protein-crystal detection with a compact multimodal multiphoton microscope.Communications biology · 2020Article
- Rapid screening ofJournal of applied crystallography · 2020Article
- Construction of gateway-compatible baculovirus expression vectors for high-throughput protein expression and in vivo microcrystal screening.Scientific reports · 2020Article
- Enhanced X-ray diffraction of in vivo-grown μNS crystals by viscous jets at XFELs.Acta crystallographica. Section F, Structural biology communications · 2020Article
- Engineering a Genetically Encoded Magnetic Protein Crystal.Nano letters · 2019Article
- Article
- Strategies for sample delivery for femtosecond crystallography.Acta crystallographica. Section D, Structural biology · 2019Review
- Towards in cellulo virus crystallography.Scientific reports · 2018Article
- Microcrystallography of Protein Crystals and In Cellulo Diffraction.Journal of visualized experiments : JoVE · 2017Article
- Review
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
While structure determination from micrometre-sized crystals used to represent a challenge, serial X-ray crystallography on microfocus beamlines at synchrotron and free-electron laser facilities greatly facilitates this process today for microcrystals and nanocrystals. In addition to typical microcrystals of purified recombinant protein, these advances have enabled the analysis of microcrystals produced inside living cells. Here, a pipeline where crystals are grown in insect cells, sorted by flow cytometry and directly analysed by X-ray diffraction is presented and applied to in vivo-grown crystals of the recombinant CPV1 polyhedrin. When compared with the analysis of purified crystals, in cellulo diffraction produces data of better quality and a gain of ∼0.35 Å in resolution for comparable beamtime usage. Importantly, crystals within cells are readily derivatized with gold and iodine compounds through the cellular membrane. Using the multiple isomorphous replacement method, a near-complete model was autobuilt from 2.7 Å resolution data. Thus, in favourable cases, an in cellulo pipeline can replace the complete workflow of structure determination without compromising the quality of the resulting model. In addition to its efficiency, this approach maintains the protein in a cellular context throughout the analysis, which reduces the risk of disrupting transient or labile interactions in protein-protein or protein-ligand complexes.
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