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Combining crystallogenesis methods to produce diffraction‐quality crystals of a psychrophilic tRNA‐maturation enzyme
Author(s) -
de Wijn Raphaël,
Hennig Oliver,
Ernst Felix G. M.,
Lorber Bernard,
Betat Heike,
Mörl Mario,
Sauter Claude
Publication year - 2018
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x18014590
Subject(s) - psychrophile , crystallization , crystal (programming language) , enzyme , trace (psycholinguistics) , biology , computational biology , chemistry , biochemistry , computer science , linguistics , philosophy , organic chemistry , programming language
The determination of conditions for the reproducible growth of well diffracting crystals is a critical step in every biocrystallographic study. On the occasion of a new structural biology project, several advanced crystallogenesis approaches were tested in order to increase the success rate of crystallization. These methods included screening by microseed matrix screening, optimization by counter‐diffusion and crystal detection by trace fluorescent labeling, and are easily accessible to any laboratory. Their combination proved to be particularly efficient in the case of the target, a 48 kDa CCA‐adding enzyme from the psychrophilic bacterium Planococcus halocryophilus . A workflow summarizes the overall strategy, which led to the production of crystals that diffracted to better than 2 Å resolution and may be of general interest for a variety of applications.