Structure of the Disulfide Bond Generating Membrane Protein DsbB in the Lipid Bilayer

The integral membrane protein DsbB in Escherichia coli is responsible for oxidizing the periplasmic protein DsbA, which forms disulfide bonds in substrate proteins. We have developed a high-resolution structural model by combining experimental X-ray and solid-state NMR with molecular dynamics (MD) s...

Full description

Saved in:
Bibliographic Details
Published in:Journal of molecular biology 2013-05, Vol.425 (10), p.1670-1682
Main Authors: Tang, Ming, Nesbitt, Anna E., Sperling, Lindsay J., Berthold, Deborah A., Schwieters, Charles D., Gennis, Robert B., Rienstra, Chad M.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacterial Proteins - biosynthesis
Bacterial Proteins - chemistry
Crystallography, X-Ray
disulfide bond generation
disulfide bonds
Disulfides - chemistry
Disulfides - metabolism
DsbB
Escherichia coli
Escherichia coli Proteins - biosynthesis
Escherichia coli Proteins - chemistry
Hydrogen Bonding
Lipid Bilayers - chemistry
Lipid Bilayers - metabolism
membrane protein
membrane proteins
Membrane Proteins - biosynthesis
Membrane Proteins - chemistry
Models, Molecular
molecular dynamics
Molecular Dynamics Simulation
Molecular Sequence Data
nuclear magnetic resonance spectroscopy
Nuclear Magnetic Resonance, Biomolecular
solid-state NMR
Substrate Specificity
topology
X-radiation
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The integral membrane protein DsbB in Escherichia coli is responsible for oxidizing the periplasmic protein DsbA, which forms disulfide bonds in substrate proteins. We have developed a high-resolution structural model by combining experimental X-ray and solid-state NMR with molecular dynamics (MD) simulations. We embedded the high-resolution DsbB structure, derived from the joint calculation with X-ray reflections and solid-state NMR restraints, into the lipid bilayer and performed MD simulations to provide a mechanistic view of DsbB function in the membrane. Further, we revealed the membrane topology of DsbB by selective proton spin diffusion experiments, which directly probe the correlations of DsbB with water and lipid acyl chains. NMR data also support the model of a flexible periplasmic loop and an interhelical hydrogen bond between Glu26 and Tyr153. [Display omitted] ► The integral membrane protein DsbB in E. coli oxidizes the periplasmic protein DsbA. ► A high-resolution structure of DsbB in the lipid bilayer is obtained. ► Selective proton spin diffusion experiments reveal membrane topology of DsbB. ► MD simulations show a flexible periplasmic loop. ► Solid-state NMR confirms an interhelical hydrogen bond between Glu26 and Tyr153.
ISSN:0022-2836
1089-8638
DOI:10.1016/j.jmb.2013.02.009