Evaluation of the data‐collection strategy for room‐temperature micro‐crystallography studied by serial synchrotron rotation crystallography combined with the humid air and glue‐coating method. Issue 3 (1st March 2021)

• Record Type: Journal Article
• Authors: Hasegawa, Kazuya; Baba, Seiki; Kawamura, Takashi; Yamamoto, Masaki; Kumasaka, Takashi
• Is Part Of: Acta crystallographica. Volume 77:Issue 3(2021)
• Page Start: 300
• Page End: 312
• Publication Date: 2021-03-01
• Subjects: serial synchrotron crystallography -- room‐temperature data collection -- humidity control -- radiation damage -- macromolecular crystallography; X-ray crystallography -- Periodicals; Crystallography -- Periodicals; Molecular biology -- Periodicals; Molecular structure -- Periodicals; Biomolecules -- Structure -- Periodicals; Cytology -- Periodicals; Biomolecules -- Structure; Crystallography; Cytology; Molecular biology; Molecular structure; X-ray crystallography; Periodicals; 548
• DOI: 10.1107/S2059798321001686
• Languages: English
• ISSNs: 2059-7983

Abstract

Abstract : A new room‐temperature (RT) data‐collection method for microcrystals was developed by combining serial synchrotron rotation crystallography with the humid air and glue‐coating method. The RT data‐collection strategy for micro‐crystallography was evaluated by examining the efficiency, the influence of non‐isomorphism and radiation damage, and the effectiveness of increasing the number of merged images. An equation was proposed that relates the achievable resolution to the total photon flux used to obtain a data set. Abstract : Synchrotron serial crystallography (SSX) is an emerging data‐collection method for micro‐crystallography on synchrotron macromolecular (MX) crystallography beamlines. At SPring‐8, the feasibility of the fixed‐target approach was examined by collecting data using a 2D raster scan combined with goniometer rotation. Results at cryogenic temperatures demonstrated that rotation is effective for efficient data collection in SSX and the method was named serial synchrotron rotation crystallography (SS‐ROX). To use this method for room‐temperature (RT) data collection, a humid air and glue‐coating (HAG) method was developed in which data were collected from polyvinyl alcohol‐coated microcrystals fixed on a loop under humidity‐controlled air. The performance and the RT data‐collection strategy for micro‐crystallography were evaluated using microcrystals of lysozyme. Although a change in unit‐cell dimensions of up to 1% was observed during data collection, the impact on data quality was marginal. A comparison of data obtained at various absorbed doses revealed that absorbed doses of up to 210 kGy were tolerable in both global and local damage. Although this limits the number of photons deposited on each crystal, increasing the number of merged images improved the resolution. On the basis of these results, an equation was proposed that relates the achievable resolution to the total photon flux used to obtain a data set.