Bacterial flagellar capping proteins adopt diverse oligomeric states.
Flagella are crucial for bacterial motility and pathogenesis. The flagellar capping protein (FliD) regulates filament assembly by chaperoning and sorting flagellin (FliC) proteins after they traverse the hollow filament and exit the growing flagellum tip. In the absence of FliD, flagella are not formed, resulting in impaired motility and infectivity. Here, we report the 2.2 Å resolution X-ray crystal structure of FliD fromPseudomonas aeruginosa, the first high-resolution structure of any FliD protein from any bacterium. Using this evidence in combination with a multitude of biophysical and functional analyses, we find thatPseudomonasFliD exhibits unexpected structural similarity to other flagellar proteins at the domain level, adopts a unique hexameric oligomeric state, and depends on flexible determinants for oligomerization. Considering that the flagellin filaments on which FliD oligomers are affixed vary in protofilament number between bacteria, our results suggest that FliD oligomer stoichiometries vary across bacteria to complement their filament assemblies.
Institute of Human Virology, University of Maryland School of Medicine, Baltimore, United States.
Sandra
0000-0002-6717-1870
S
Postel
Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, United States.
Daniel
D
Deredge
Institute of Human Virology, University of Maryland School of Medicine, Baltimore, United States.
Daniel A
DA
Bonsor
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, United States.
Xiong
X
Yu
Department of Biology, University of Konstanz, Konstanz, Germany.
Kay
K
Diederichs
Department of Biotechnology, Institute of Biochemistry, Biotechnology and Bioinformatics, Technische Universität Braunschweig, Braunschweig, Germany.
Saskia
S
Helmsing
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel.
Aviv
A
Vromen
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel.
Assaf
A
Friedler
Department of Biotechnology, Institute of Biochemistry, Biotechnology and Bioinformatics, Technische Universität Braunschweig, Braunschweig, Germany.
Michael
M
Hust
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, United States.
Edward H
0000-0003-4844-5212
EH
Egelman
Department of Chemistry and Biochemistry, University of Maryland College Park, Baltimore, United States.
Dorothy
D
Beckett
Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, United States.
Patrick L
PL
Wintrode
Institute of Human Virology, University of Maryland School of Medicine, Baltimore, United States.|Department of Medicine, University of Maryland School of Medicine, Baltimore, United States.|Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, United States.
Eric J
0000-0003-0478-3033
EJ
Sundberg
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D005408:Flagellin / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D055503:Protein Multimerization; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
D001426:Bacterial Proteins; D005408:Flagellin; C076733:FlaD protein, Bacteria
Pseudomonas; X-ray crystallography; analytical ultracentrifugation; biophysics; flagella; hydrogen-deuterium exchange; infectious disease; microbiology; structural biology
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