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Title: Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams

Abstract

High-resolution structural information was obtained from lysozyme microcrystals (20 µm in the largest dimension) using raster-scanning serial protein crystallography on micro- and nano-focused beamlines at the ESRF. Data were collected at room temperature (RT) from crystals sandwiched between two silicon nitride wafers, thereby preventing their drying, while limiting background scattering and sample consumption. In order to identify crystal hits, new multi-processing and GUI-driven Python-based pre-analysis software was developed, named NanoPeakCell, that was able to read data from a variety of crystallographic image formats. Further data processing was carried out using CrystFEL, and the resultant structures were refined to 1.7 Å resolution. The data demonstrate the feasibility of RT raster-scanning serial micro- and nano-protein crystallography at synchrotrons and validate it as an alternative approach for the collection of high-resolution structural data from micro-sized crystals. Advantages of the proposed approach are its thriftiness, its handling-free nature, the reduced amount of sample required, the adjustable hit rate, the high indexing rate and the minimization of background scattering.

Authors:
 [1];  [2];  [3];  [3];  [3];  [4];  [1]
  1. Universite´ Grenoble Alpes, Genoble (France); CNRS, Grenoble (France); CEA, Genable (France)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. European Synchrotron Radiation Facility, Grenoble (France)
  4. European Synchrotron Radiation Facility, Grenoble (France); Ghent Univ., Ghent (Belgium)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1213447
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section D: Biological Crystallography (Online)
Additional Journal Information:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography (Online); Journal Volume: 71; Journal Issue: 5; Journal ID: ISSN 1399-0047
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; serial crystallography; hit finding; raster scanning; background subtraction; radiation damage

Citation Formats

Coquelle, Nicolas, Brewster, Aaron S., Kapp, Ulrike, Shilova, Anastasya, Weinhausen, Britta, Burghammer, Manfred, and Colletier, Jacques -Philippe. Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams. United States: N. p., 2015. Web. doi:10.1107/S1399004715004514.
Coquelle, Nicolas, Brewster, Aaron S., Kapp, Ulrike, Shilova, Anastasya, Weinhausen, Britta, Burghammer, Manfred, & Colletier, Jacques -Philippe. Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams. United States. doi:10.1107/S1399004715004514.
Coquelle, Nicolas, Brewster, Aaron S., Kapp, Ulrike, Shilova, Anastasya, Weinhausen, Britta, Burghammer, Manfred, and Colletier, Jacques -Philippe. Sat . "Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams". United States. doi:10.1107/S1399004715004514. https://www.osti.gov/servlets/purl/1213447.
@article{osti_1213447,
title = {Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams},
author = {Coquelle, Nicolas and Brewster, Aaron S. and Kapp, Ulrike and Shilova, Anastasya and Weinhausen, Britta and Burghammer, Manfred and Colletier, Jacques -Philippe},
abstractNote = {High-resolution structural information was obtained from lysozyme microcrystals (20 µm in the largest dimension) using raster-scanning serial protein crystallography on micro- and nano-focused beamlines at the ESRF. Data were collected at room temperature (RT) from crystals sandwiched between two silicon nitride wafers, thereby preventing their drying, while limiting background scattering and sample consumption. In order to identify crystal hits, new multi-processing and GUI-driven Python-based pre-analysis software was developed, named NanoPeakCell, that was able to read data from a variety of crystallographic image formats. Further data processing was carried out using CrystFEL, and the resultant structures were refined to 1.7 Å resolution. The data demonstrate the feasibility of RT raster-scanning serial micro- and nano-protein crystallography at synchrotrons and validate it as an alternative approach for the collection of high-resolution structural data from micro-sized crystals. Advantages of the proposed approach are its thriftiness, its handling-free nature, the reduced amount of sample required, the adjustable hit rate, the high indexing rate and the minimization of background scattering.},
doi = {10.1107/S1399004715004514},
journal = {Acta Crystallographica. Section D: Biological Crystallography (Online)},
number = 5,
volume = 71,
place = {United States},
year = {2015},
month = {4}
}

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Cited by: 33 works
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