Projects per year
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 as well as its N-terminal DNA binding domain withstand crystallization. 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 a= 317 Å, diffract to 4.0 Å resolution and likely contain four PaaR2 monomers and four nanobody monomers in their asymmetric unit. Crystals of two truncates containing the N-terminal helix-turn-helix domain also interact with Nb33 and the corresponding co-crystals diffract to 1.6 and 1.75 Å resolution.
Original language | English |
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Article number | 11 |
Pages (from-to) | 374-384 |
Number of pages | 11 |
Journal | Acta Crystallographica Section F - Structural Biology Communications |
Volume | 77 |
Issue number | 10 |
DOIs | |
Publication status | Published - 1 Oct 2021 |
Keywords
- Nanobody aided crystallography
- Toxin-Antitoxin module
- Transcription regulation
- Transcription Factor
- Structural Biology
- Crystallography
- Molecular Biophysics
Fingerprint
Dive into the research topics of 'Nanobody-aided crystallization of the transcription regulator PaaR2 from Escherichia coli O157:H7'. Together they form a unique fingerprint.Projects
- 1 Finished
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FWOAL831: Regulation of persistence by the paaAR-paaA2-parE2 and cog4197-duf1019 operons from E. coli O157
1/01/17 → 31/12/20
Project: Fundamental
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Bistable expression of a toxin-antitoxin system located in a cryptic prophage of Escherichia coliO157:H7
Jurenas, D., Fraikin, N., Goormaghtigh, F., De Bruyn, P., Vandervelde, A., Jove, T., Charlier, D., Loris, R. & Van Melderen, L., 21 Dec 2021, In: mBio. 12, 6, 15 p., e0294721.Research output: Contribution to journal › Article › peer-review
Open AccessFile3 Citations (Scopus)37 Downloads (Pure) -
Prokaryote toxin-antitoxin modules: complex regulation of an unclear function
De Bruyn, P., Girardin, Y. & Loris, R., Jun 2021, In: Protein Sci. 30, 6, p. 1103-1113 11 p.Research output: Contribution to journal › Article › peer-review
Open AccessFile9 Citations (Scopus)107 Downloads (Pure) -
Toxin-antitoxin modules: functional characterization of transcription regulation and potential use against pathogenic bacteria
De Bruyn, P., 2021, Brussels.Research output: Thesis › PhD Thesis
Activities
- 1 Member of PhD committee
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Department of Bio-engineering Sciences (Organisational unit)
Eveline Peeters (Chair), Inge Van Molle (Jury), Kim Roelants (Jury), Remy Loris (Supervisor), Henri De Greve (Supervisor), Tanel Tenson (Jury) & Jan Michiels (Jury)
2 Sep 2021Activity: Membership › Member of PhD committee