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Room-temperature serial crystallography using a kinetically optimized microfluidic device for protein crystallization and on-chip X-ray diffraction

Authors :
Michael Heymann
Achini Opthalage
Jennifer L. Wierman
Sathish Akella
Doletha M. E. Szebenyi
Sol M. Gruner
Seth Fraden
Source :
IUCrJ, Vol 1, Iss 5, Pp 349-360 (2014)
Publication Year :
2014
Publisher :
International Union of Crystallography, 2014.

Abstract

An emulsion-based serial crystallographic technology has been developed, in which nanolitre-sized droplets of protein solution are encapsulated in oil and stabilized by surfactant. Once the first crystal in a drop is nucleated, the small volume generates a negative feedback mechanism that lowers the supersaturation. This mechanism is exploited to produce one crystal per drop. Diffraction data are measured, one crystal at a time, from a series of room-temperature crystals stored on an X-ray semi-transparent microfluidic chip, and a 93% complete data set is obtained by merging single diffraction frames taken from different unoriented crystals. As proof of concept, the structure of glucose isomerase was solved to 2.1 Å, demonstrating the feasibility of high-throughput serial X-ray crystallography using synchrotron radiation.

Details

Language :
English
ISSN :
20522525
Volume :
1
Issue :
5
Database :
Directory of Open Access Journals
Journal :
IUCrJ
Publication Type :
Academic Journal
Accession number :
edsdoj.00cb9c7e57fe4ff5b510ab2e9bd56616
Document Type :
article
Full Text :
https://doi.org/10.1107/S2052252514016960