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Production, biophysical characterization and crystallization of Pseudomonas putida GraA and its complexes with GraT and the graTA operator
Author(s) -
Talavera Ariel,
Tamman Hedvig,
Ainelo Andres,
Hadži San,
Garcia-Pino Abel,
Hõrak Rita,
Konijnenberg Albert,
Loris Remy
Publication year - 2017
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/s2053230x17009438
Subject(s) - pseudomonas putida , crystallization , resolution (logic) , operon , crystallography , chemistry , stereochemistry , escherichia coli , biochemistry , organic chemistry , enzyme , artificial intelligence , computer science , gene
The graTA operon from Pseudomonas putida encodes a toxin–antitoxin module with an unusually moderate toxin. Here, the production, SAXS analysis and crystallization of the antitoxin GraA, the GraTA complex and the complex of GraA with a 33 bp operator fragment are reported. GraA forms a homodimer in solution and crystallizes in space group P 2 1 , with unit‐cell parameters a = 66.9, b = 48.9, c = 62.7 Å, β = 92.6°. The crystals are likely to contain two GraA dimers in the asymmetric unit and diffract to 1.9 Å resolution. The GraTA complex forms a heterotetramer in solution. Crystals of the GraTA complex diffracted to 2.2 Å resolution and are most likely to contain a single heterotetrameric GraTA complex in the asymmetric unit. They belong to space group P 4 1 or P 4 3 , with unit‐cell parameters a = b = 56.0, c = 128.2 Å. The GraA–operator complex consists of a 33 bp operator region that binds two GraA dimers. It crystallizes in space group P 3 1 or P 3 2 , with unit‐cell parameters a = b = 105.6, c = 149.9 Å. These crystals diffract to 3.8 Å resolution.