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

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 , 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 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.

Source: Acta Crystallographica Section F - July 26, 2017 Category: Biochemistry Authors: Talavera, A. Tamman, H. Ainelo, A. Had ž i, S. Garcia-Pino, A. H õ rak, R. Konijnenberg, A. Loris, R. Tags: persistence toxin – antitoxin module protein DNA complex macromolecular complex GraT GraA Pseudomonas putida ribosome biogenesis research communications Source Type: research