De Bruyn, P, Prolič-Kalinšek, M, Vandervelde, A et al. (7 more authors) (2021) Nanobody-aided crystallization of the transcription regulator PaaR2 from Escherichia coli O157:H7. Acta Crystallographica Section F: Structural Biology Communications, 77 (10). pp. 374-384. ISSN 2053-230X
Abstract
paaR2–paaA2–parE2 is a three-component toxin–antitoxin module found in prophage CP-993P of Escherichia coli O157:H7. Transcription regulation of this module occurs via the 123-amino-acid regulator PaaR2, which forms a large oligomeric structure. Despite appearing to be well folded, PaaR2 withstands crystallization, as does its N-terminal DNA-binding domain. Native mass spectrometry was used to screen for nanobodies that form a unique complex and stabilize the octameric structure of PaaR2. One such nanobody, Nb33, allowed crystallization of the protein. The resulting crystals belong to space group F432, with unit-cell parameter a = 317 Å, diffract to 4.0 Å resolution and are likely to contain four PaaR2 monomers and four nanobody monomers in the asymmetric unit. Crystals of two truncates containing the N-terminal helix–turn–helix domain also interact with Nb33, and the corresponding co-crystals diffracted to 1.6 and 1.75 Å resolution.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2021 International Union of Crystallography. Reproduced in accordance with the publisher's self-archiving policy. |
Keywords: | toxin–antitoxin modules; transcription regulation; macromolecular complexes; nanobody-aided crystallization; crystallization chaperones; Escherichia coli O157:H7; PaaR2 |
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: | 24 Nov 2021 15:42 |
Last Modified: | 25 Jun 2023 22:49 |
Status: | Published |
Publisher: | International Union of Crystallography |
Identification Number: | 10.1107/s2053230x21009006 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:180323 |