Garcia-Rodriguez, G, Talavera Perez, A, Konijnenberg, A et al. (3 more authors) (2020) The Escherichia coli RnlA–RnlB toxin–antitoxin complex: production, characterization and crystallization. Acta Crystallographica Section F Structural Biology Communications, 76 (1). pp. 31-39. ISSN 2053-230X
Abstract
The Escherichia coli rnlAB operon encodes a toxin–antitoxin module that is involved in protection against infection by bacteriophage T4. The full-length RnlA–RnlB toxin–antitoxin complex as well as the toxin RnlA were purified to homogeneity and crystallized. When the affinity tag is placed on RnlA, RnlB is largely lost during purification and the resulting crystals exclusively comprise RnlA. A homogeneous preparation of RnlA–RnlB containing stoichiometric amounts of both proteins could only be obtained using a His tag placed C-terminal to RnlB. Native mass spectrometry and SAXS indicate a 1:1 stoichiometry for this RnlA–RnlB complex. Crystals of the RnlA–RnlB complex belonged to space group C2, with unit-cell parameters a = 243.32, b = 133.58, c = 55.64 Å, β = 95.11°, and diffracted to 2.6 Å resolution. The presence of both proteins in the crystals was confirmed and the asymmetric unit is likely to contain a heterotetramer with RnlA2:RnlB2 stoichiometry.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2020 International Union of Crystallography. Reproduced in accordance with the publisher's self-archiving policy. |
Keywords: | toxin–antitoxin; bacterial stress response; macromolecular complex; Escherichia coli |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 06 Apr 2020 10:26 |
Last Modified: | 25 Jun 2023 22:13 |
Status: | Published |
Publisher: | International Union of Crystallography |
Identification Number: | 10.1107/s2053230x19017175 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:159003 |