Projects per year
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 P2 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 P4 1 or P4 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 P3 1 or P3 2, with unit-cell parameters a = b = 105.6, c = 149.9 Å. These crystals diffract to 3.8 Å resolution.The antitoxin GraA from P. putida and its complexes with the toxin GraT and with the 33 bp operator of the graTA operon were crystallized.
Original language | English |
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Pages (from-to) | 455-462 |
Number of pages | 8 |
Journal | Acta Crystallographica Section F - Structural Biology Communications |
Volume | F73 |
Issue number | 8 |
DOIs | |
Publication status | Published - Aug 2017 |
Keywords
- Amino Acid Sequence
- Antitoxins/chemistry
- Bacterial Proteins/chemistry
- Bacterial Toxins/chemistry
- Cloning, Molecular
- Crystallization
- Crystallography, X-Ray
- Escherichia coli/genetics
- Gene Expression
- Gene Expression Regulation, Bacterial
- Genetic Vectors/chemistry
- Operon
- Protein Binding
- Protein Multimerization
- Pseudomonas putida/chemistry
- Recombinant Fusion Proteins/chemistry
- X-Ray Diffraction
Fingerprint
Dive into the research topics of 'Production, biophysical characterization and crystallization of Pseudomonas putida GraA and its complexes with GraT and the graTA operator'. Together they form a unique fingerprint.Projects
- 4 Finished
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FWOAL756: Regulatory mechanisms in bacterial toxin-antitoxin modules
1/01/15 → 31/12/18
Project: Fundamental
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FWOAL699: ESRF and DUBBLE: Synchrotron X-rays for revealing the structure and function of molecules and materials
Loris, R., Remaut, H. K., Goderis, B., Janssens, K., Kirschhock, C. E. A., Van Bael, M., Poelman, D., Temst, K., Adriaens, A. & Laszlo, V.
1/01/13 → 31/12/17
Project: Fundamental
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HERC20: Structural biology of large and flexible macromolecules in solution: Small angle X-Ray Scatter (Bio-SAXS) and Multi-angle light scatter (MALS) (UABR/11/012)
Loris, R., Steyaert, J., Versées, W., Van Nuland, N., Remaut, H. K., Tompa, P., Savvides, S. & De Veylder, L.
19/11/12 → 24/04/17
Project: Fundamental
Activities
- 1 Talk or presentation at a conference
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12th Meeting of the Slovenian Biochemical Society with international participation
Remy Loris (Speaker)
20 Sept 2017 → 23 Sept 2017Activity: Talk or presentation › Talk or presentation at a conference