Computational and biochemical analysis of type IV Pilus dynamics and stability

Yasaman Karami (1) , Aracelys López-Castilla (1, 2) , Andrea Ori (3) , Jenny-Lee Thomassin (3) , Benjamin Bardiaux (1) , Therese Malliavin (1) , Nadia Izadi-Pruneyre (1, 2) , Olivera Francetic (3) , Michael Nilges (1)

1 Bioinformatique structurale - Structural Bioinformatics
2 Résonance Magnétique Nucléaire des Biomolécules
3 Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions

Type IV pili (T4P) are distinctive dynamic filaments at the surface of many bacteria that can rapidly extend, retract and withstand strong forces. T4P are important virulence factors in many human pathogens, including Enterohemorrhagic Escherichia coli (EHEC). The structure of the EHEC T4P has been determined by integrating Nuclear Magnetic Resonance (NMR) and cryo-electron microscopy data. To better understand pilus assembly, stability and function, we performed a total of 108 µs all-atom molecular dynamics simulations of wild-type and mutant T4P. Extensive characterization of the conformational landscape of T4P in different conditions of temperature, pH and ionic strength was complemented by targeted mutagenesis and biochemical analyses. Our simulations and NMR experiments revealed a conserved set of residues defining a novel calcium-binding site at the interface between three pilin subunits. Calcium binding enhanced T4P stability ex vivo and in vitro, supporting the role of this binding site as a potential pocket for drug design.

Domaines: Biologie moléculaire

Yasaman Karami, Aracelys López-Castilla, Andrea Ori, Jenny-Lee Thomassin, Benjamin Bardiaux, et al.. Computational and biochemical analysis of type IV Pilus dynamics and stability. 2021. ⟨pasteur-03380000v1⟩