Characterization of H/D exchange in type 1 pili by proton-detected solid-state NMR and molecular dynamics simulations.
J Biomol NMR
; 73(6-7): 281-291, 2019 Jul.
Article
in En
| MEDLINE
| ID: mdl-31028572
ABSTRACT
Uropathogenic Escherichia coli invades and colonizes hosts by attaching to cells using adhesive pili on the bacterial surface. Although many biophysical techniques have been used to study the structure and mechanical properties of pili, many important details are still unknown. Here we use proton-detected solid-state NMR experiments to investigate solvent accessibility and structural dynamics. Deuterium back-exchange at labile sites of the perdeuterated, fully proton back-exchanged pili was conducted to investigate hydrogen/deuterium (H/D) exchange patterns of backbone amide protons in pre-assembled pili. We found distinct H/D exchange patterns in lateral and axial intermolecular interfaces in pili. Amide protons protected from H/D exchange in pili are mainly located in the core region of the monomeric subunit and in the lateral intermolecular interface, whereas the axial intermolecular interface and the exterior region of pili are highly exposed to H/D exchange. Additionally, we performed molecular dynamics simulations of the type 1 pilus rod and estimated the probability of H/D exchange based on hydrogen bond dynamics. The comparison of the experimental observables and simulation data provides insights into stability and mechanical properties of pili.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protons
/
Nuclear Magnetic Resonance, Biomolecular
/
Fimbriae Proteins
/
Deuterium
/
Molecular Dynamics Simulation
/
Hydrogen
Language:
En
Journal:
J Biomol NMR
Journal subject:
BIOLOGIA MOLECULAR
/
DIAGNOSTICO POR IMAGEM
/
MEDICINA NUCLEAR
Year:
2019
Document type:
Article
Affiliation country:
Germany