On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

Lieske, Julia; Cerv, Maximilian; Kreida, Stefan; Komadina, Dana; Fischer, Janine; Barthelmess, Miriam; Fischer, Pontus; Pakendorf, Tim; Yefanov, Oleksandr; Mariani, Valerio; Seine, Thomas; Ross, Breyan H.; Crosas, Eva; Lorbeer, Olga; Burkhardt, Anja; Lane, Thomas J.; Guenther, Sebastian; Bergtholdt, Julian; Schoen, Silvan; Törnroth-Horsefield, Susanna; Chapman, Henry N.; Meents, Alke

doi:10.1107/s2052252519007395

Title: On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

Journal Article · 19 June 2019 · IUCrJ

DOI: https://doi.org/10.1107/s2052252519007395 · OSTI ID:1617994

^[1]; Cerv, Maximilian ^[1]; Kreida, Stefan ^[2]; Komadina, Dana ^[1]; Fischer, Janine ^[1]; Barthelmess, Miriam ^[1]; Fischer, Pontus ^[1]; Pakendorf, Tim ^[1]; Yefanov, Oleksandr ^[1];

^[1]; Seine, Thomas ^[3];

^[4]; Crosas, Eva ^[1]; Lorbeer, Olga ^[1];

^[1]; Lane, Thomas J. ^[5]; Guenther, Sebastian ^[5]; Bergtholdt, Julian ^[1]; Schoen, Silvan ^[1]; Törnroth-Horsefield, Susanna ^[2] more »

Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
Lund Univ. (Sweden)
Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); EMBL, Hamburg (Germany)
Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Max Planck Institute of Biochemistry (Germany)
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg (Germany)

Efficient and reliable sample delivery has remained one of the bottlenecks for serial crystallography experiments. Compared with other methods, fixed-target sample delivery offers the advantage of significantly reduced sample consumption and shorter data collection times owing to higher hit rates. Here, a new method of on-chip crystallization is reported which allows the efficient and reproducible growth of large numbers of protein crystals directly on micro-patterned silicon chips forin-situserial crystallography experiments. Crystals are grown by sitting-drop vapor diffusion and previously established crystallization conditions can be directly applied. By reducing the number of crystal-handling steps, the method is particularly well suited for sensitive crystal systems. Excessive mother liquor can be efficiently removed from the crystals by blotting, and no sealing of the fixed-target sample holders is required to prevent the crystals from dehydrating. As a consequence, `naked' crystals are obtained on the chip, resulting in very low background scattering levels and making the crystals highly accessible for external manipulation such as the application of ligand solutions. Serial diffraction experiments carried out at cryogenic temperatures at a synchrotron and at room temperature at an X-ray free-electron laser yielded high-quality X-ray structures of the human membrane protein aquaporin 2 and two new ligand-bound structures of thermolysin and the human kinase DRAK2. The results highlight the applicability of the method for future high-throughput on-chip screening of pharmaceutical compounds.

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Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE

Grant/Contract Number:: AC02-76SF00515; 654220; 609920; 05K2018 – 2017-06727 MXD; 2010-5208; 2012-2849

OSTI ID:: 1617994

Alternate ID(s):: OSTI ID: 1546938

Journal Information:: IUCrJ, Vol. 6, Issue 4; ISSN 2052-2525

Publisher:: International Union of CrystallographyCopyright Statement

Country of Publication:: United States

Language:: English

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A fixed-target platform for serial femtosecond crystallography in a hydrated environment Shelby, M. L.; Gilbile, D.; Grant, T. D. IUCrJ, Vol. 7, Issue 1 https://doi.org/10.1107/s2052252519014003	journal	January 2020
A fixed-target platform for serial femtosecond crystallography in a hydrated environment Shelby, M. L.; Gilbile, D.; Grant, T. D. Deutsches Elektronen-Synchrotron, DESY, Hamburg https://doi.org/10.3204/pubdb-2019-05316	text	January 2020

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
serial crystallography
silicon chip
fixed-target crystallography
in-situ diffraction
vapor diffusion
ligand binding
ligand soaking
drug discovery
protein structure
X-ray crystallography

Title: On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

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