Solid-state NMR and molecular dynamics simulations reveal the oligomeric ion-channels of TM2-GABA(A) stabilized by intermolecular hydrogen bonding.
The second transmembrane (TM2) domain of GABA(A) receptor forms the inner-lining surface of chloride ion-channel and plays important roles in the function of the receptor protein. In this study, we report the first structure of TM2 in lipid bilayers determined using solid-state NMR and MD simulations. The interatomic (13)C-(15)N distances measured from REDOR magic angle spinning experiments on multilamellar vesicles, containing a TM2 peptide site specifically labeled with (13)C' and (15)N isotopes, were used to determine the secondary structure of the peptide. The (15)N chemical shift and (1)H-(15)N dipolar coupling parameters measured from PISEMA experiments on mechanically aligned phospholipid bilayers, containing a TM2 peptide site specifically labeled with (15)N isotopes, under static conditions were used to determine the membrane orientation of the peptide. Our results reveal that the TM2 peptide forms an alpha helical conformation with a tilted transmembrane orientation, which is unstable as a monomer but stable as pentameric oligomers as indicated by MD simulations. Even though the peptide consists of a number of hydrophilic residues, the transmembrane folding of the peptide is stabilized by intermolecular hydrogen bondings between the side chains of Ser and Thr residues as revealed by MD simulations. The results also suggest that peptide-peptide interactions in the tilted transmembrane orientation overcome the hydrophobic mismatch between the peptide and bilayer thickness.
Kandasamy
Senthil K
SK
Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-1055, USA.
Lee
Dong-Kuk
DK
Nanga
Ravi P R
RP
Xu
Jiadi
J
Santos
Jose S
JS
Larson
Ronald G
RG
Ramamoorthy
Ayyalusamy
A
D018118:Chloride Channels / Q000737:chemistry / Q000378:metabolism; D003198:Computer Simulation; D006860:Hydrogen Bonding; D008051:Lipid Bilayers; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D017433:Protein Structure, Secondary; D011963:Receptors, GABA-A / Q000737:chemistry / Q000502:physiology*
D018118:Chloride Channels; D008051:Lipid Bilayers; D011963:Receptors, GABA-A