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Lookup NU author(s): Adam Crawshaw, Dr Arnaud Basle, Professor Paula SalgadoORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© Crawshaw et al. 2020.Many biologists are now routinely seeking to determine the three-dimensional structures of their proteins of choice, illustrating the importance of this knowledge, but also of the simplification and streamlining of structure-determination processes. Despite the fact that most software packages offer simple pipelines, for the non-expert navigating the outputs and understanding the key aspects can be daunting. Here, the structure determination of the type IV pili (TFP) protein PilA1 from Clostridioides difficile is used to illustrate the different steps involved, the key decision criteria and important considerations when using the most common pipelines and software. Molecular-replacement pipelines within CCP4i2 are presented to illustrate the more commonly used processes. Previous knowledge of the biology and structure of TFP pilins, particularly the presence of a long, N-terminal α-helix required for pilus formation, allowed informed decisions to be made during the structure-determination strategy. The PilA1 structure was finally successfully determined using ARCIMBOLDO and the ab initio MR strategy used is described.
Author(s): Crawshaw AD, Basle A, Salgado PS
Publication type: Article
Publication status: Published
Journal: Acta Crystallographica Section D: Structural Biology
Year: 2020
Volume: 76
Pages: 261-271
Online publication date: 01/03/2020
Acceptance date: 14/01/2020
Date deposited: 23/03/2020
ISSN (electronic): 2059-7983
Publisher: International Union of Crystallography
URL: https://doi.org/10.1107/S2059798320000467
DOI: 10.1107/S2059798320000467
PubMed id: 32133990
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