Hybrid Structure of the Type 1 Pilus of Uropathogenic Escherichia coli.
Type 1 pili are filamentous protein assemblies on the surface of Gram-negative bacteria that mediate adhesion to host cells during the infection process. The molecular structure of type 1 pili remains elusive on the atomic scale owing to their insolubility and noncrystallinity. Herein we describe an approach for hybrid-structure determination that is based on data from solution-state NMR spectroscopy on the soluble subunit and solid-state NMR spectroscopy and STEM data on the assembled pilus. Our approach is based on iterative modeling driven by structural information extracted from different sources and provides a general tool to access pseudo atomic structures of protein assemblies with complex subunit folds. By using this methodology, we determined the local conformation of the FimA pilus subunit in the context of the assembled type 1 pilus, determined the exact helical pilus architecture, and elucidated the intermolecular interfaces contributing to pilus assembly and stability with atomic detail.
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).|Université de Bordeaux/CBMN UMR5248, Institut Européen de Chimie et Biologie (IECB), 2 rue Robert Escarpit, 33600 Pessac (France).
Birgit
B
Habenstein
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).|Université de Bordeaux/CBMN UMR5248, Institut Européen de Chimie et Biologie (IECB), 2 rue Robert Escarpit, 33600 Pessac (France).
Antoine
A
Loquet
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).|Institut für Biologie, Humboldt-Universität zu Berlin, Invalidenstrasse 110, 10115 Berlin (Germany).|Department of Molecular Biophysics, Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany).
Songhwan
S
Hwang
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).
Karin
K
Giller
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).
Suresh Kumar
SK
Vasa
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany).
Stefan
S
Becker
Felix Bernstein Institute for Mathematical Statistics in the Biosciences, University of Göttingen, Goldschmidtstrasse 7, 37077 Göttingen (Germany). mhabeck@gwdg.de.|Statistical Inverse Problems in Biophysics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany). mhabeck@gwdg.de.
Michael
M
Habeck
Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany). alange@fmp-berlin.de.|Institut für Biologie, Humboldt-Universität zu Berlin, Invalidenstrasse 110, 10115 Berlin (Germany). alange@fmp-berlin.de.|Department of Molecular Biophysics, Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany). alange@fmp-berlin.de.
Adam
A
Lange
D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry; D056689:Uropathogenic Escherichia coli / Q000737:chemistry* / Q000648:ultrastructure
D021122:Protein Subunits
filamentous protein assemblies; iterative modeling; pilus structure; solid-state NMR spectroscopy; structure elucidation