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Bacterial Filamentous Appendages Investigated by Solid-State NMR Spectroscopy

  • Birgit Habenstein
  • Antoine Loquet
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1615)

Abstract

The assembly of filamentous appendages at the surface of bacteria is essential in many infection mechanisms. The extent of mechanical, dynamical, and functional properties of such appendages is very diverse, ranging from a structural scaffold of the pathogen–host cell interaction to cell motility, surface adhesion, or the export of virulence effectors. In particular, the architectures of several bacterial secretion systems have revealed the presence of filamentous architectures, known as pili, fimbriae, andneedles. At the macroscopic level, filamentous bacterial appendages appear as thin extracellular filaments of several nanometers in diameter and up to several microns in length. The structural characterization of these appendages at atomic-scale resolution represents an extremely challenging task because of their inherent noncrystallinity and very poor solubility. Here, we describe protocols based on recent advances in solid-state NMR spectroscopy to investigate the secondary structure, subunit–subunit protein interactions, symmetry parameters, and atomic architecture of bacterial filaments.

Key words

Solid-state nuclear magnetic resonance Structure determination Pilus Needle Protein assembly Protein complex Helical symmetry 

Notes

Acknowledgments

The authors thank their past and present colleagues, in particular Prof. Adam Lange at the Leibniz-Institut für Molekulare Pharmakologie for his guidance during the author postdoctoral periods and his main intellectual contribution for the T3SS needle and type I pilus projects. This work was further supported by the Fondation pour la Recherche Médicale (FRM-AJE20140630090 to A.L.), the ANR (13-PDOC-0017-01 to B.H. and ANR-14-CE09-0020-01 to A.L.), the FP7 program (FP7-PEOPLE-2013-CIG to A.L.), the IdEx Bordeaux University (Chaire d’Installation to B.H.)and the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC Starting Grant to A.L., agreement 105945). Erick Dufourc is acknowledged for his continuous support.

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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR5248 CBMN), CNRSUniversity of Bordeaux, Institut Européen de Chimie et BiologiePessacFrance

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