Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi

Halina Mikolajek, Juan Sanchez-Weatherby, James Sandy, Richard J. Gildea, Ivan Campeotto, Harish Cheruvara, John D. Clarke, Toshana Foster, Sotaro Fujii, Ian T. Paulsen, Bhumika S. Shah, Michael Hough*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)
45 Downloads (Pure)

Abstract

The utility of X-ray crystal structures determined under ambient-temperature conditions is becoming increasingly recognized. Such experiments can allow protein dynamics to be characterized and are particularly well suited to challenging protein targets that may form fragile crystals that are difficult to cryo-cool. Room-temperature data collection also enables time-resolved experiments. In contrast to the high-throughput highly automated pipelines for determination of structures at cryogenic temperatures widely available at synchrotron beamlines, room-temperature methodology is less mature. Here, the current status of the fully automated ambient-temperature beamline VMXi at Diamond Light Source is described, and a highly efficient pipeline from protein sample to final multi-crystal data analysis and structure determination is shown. The capability of the pipeline is illustrated using a range of user case studies representing different challenges, and from high and lower symmetry space groups and varied crystal sizes. It is also demonstrated that very rapid structure determination from crystals in situ within crystallization plates is now routine with minimal user intervention.
Original languageEnglish
Pages (from-to)420-429
Number of pages10
JournalIUCrJ
Volume10
Issue numberPart 4
Early online date19 May 2023
DOIs
Publication statusPublished - Jul 2023

Bibliographical note

Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • room temperature
  • in situ
  • multi-crystal
  • crystallization pipelines
  • automated data collection
  • structural biology
  • radiation damage
  • X-ray crystallography
  • VMXi

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