|
Title
|
|
|
|
Nanobody-aided crystallization of the transcription regulator PaaR2 from Escherichia coli O157:H7
|
|
|
Author
|
|
|
|
|
|
|
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. |
|
|
|
Language
|
|
|
|
English
|
|
|
Source (journal)
|
|
|
|
Acta Chrystallographica Section F. - -
|
|
|
Publication
|
|
|
|
2021
|
|
|
ISSN
|
|
|
|
2053-230X
|
|
|
DOI
|
|
|
|
10.1107/S2053230X21009006
|
|
|
Volume/pages
|
|
|
|
77
:10
(2021)
, p. 374-384
|
|
|
ISI
|
|
|
|
000711166300007
|
|
|
Pubmed ID
|
|
|
|
34605442
|
|
|
Full text (Publisher's DOI)
|
|
|
|
|
|
|
Full text (open access)
|
|
|
|
|
|
|
Full text (publisher's version - intranet only)
|
|
|
|
|
|