TY - JOUR
T1 - Structure of the disulfide bond generating membrane protein DsbB in the lipid bilayer
AU - Tang, Ming
AU - Nesbitt, Anna E.
AU - Sperling, Lindsay J.
AU - Berthold, Deborah A.
AU - Schwieters, Charles D.
AU - Gennis, Robert B.
AU - Rienstra, Chad M.
N1 - Funding Information:
This research was supported by the National Institutes of Health ( R01GM075937 , S10RR025037 , R01HL103999, and R01GM073770 ARRA supplement to C.M.R., and NRSA F32GM095344 to A.E.N), a Molecular Biophysics Training Grant ( PHS 5 T32 GM008276 ) and Ullyot Fellowship to L.J.S., and the National Institutes of Health Intramural Research Program of CIT to C.D.S. The authors thank the School of Chemical Sciences NMR Facility at the University of Illinois at Urbana-Champaign for assistance with data acquisition, Mike Hallock for helpful assistance with structure calculations, and Dr. Taras Pogorelov for helpful discussion with MD simulations.
PY - 2013/5/27
Y1 - 2013/5/27
N2 - 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.
AB - 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.
KW - DsbB
KW - disulfide bond generation
KW - membrane protein
KW - molecular dynamics simulation
KW - solid-state NMR
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U2 - 10.1016/j.jmb.2013.02.009
DO - 10.1016/j.jmb.2013.02.009
M3 - Article
C2 - 23416557
AN - SCOPUS:84877580955
SN - 0022-2836
VL - 425
SP - 1670
EP - 1682
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 10
ER -