Production, biophysical characterization and crystallization of Pseudomonas putida GraA and its complexes with GraT and the graTA operator.

ABSTRACT: 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 P21, with unit-cell parameters a = 66.9, b = 48.9, c = 62.7?Å, ? = 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 P41 or P43, 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 P31 or P32, with unit-cell parameters a = b = 105.6, c = 149.9?Å. These crystals diffract to 3.8?Å resolution.

SUBMITTER: Talavera A 

PROVIDER: S-EPMC5544002 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature