Investigating structural changes in the lipid bilayer upon insertion of the transmembrane domain of the membrane-bound protein phospholamban utilizing 31P and 2H solid-state NMR spectroscopy.
Phospholamban (PLB) is a 52-amino acid integral membrane protein that regulates the flow of Ca(2+) ions in cardiac muscle cells. In the present study, the transmembrane domain of PLB (24-52) was incorporated into phospholipid bilayers prepared from 1-palmitoyl-2-oleoyl-sn-glycero-phosphocholine (POPC). Solid-state (31)P and (2)H NMR experiments were carried out to study the behavior of POPC bilayers in the presence of the hydrophobic peptide PLB at temperatures ranging from 30 degrees C to 60 degrees C. The PLB peptide concentration varied from 0 mol % to 6 mol % with respect to POPC. Solid-state (31)P NMR spectroscopy is a valuable technique to study the different phases formed by phospholipid membranes. (31)P NMR results suggest that the transmembrane protein phospholamban is incorporated successfully into the bilayer and the effects are observed in the lipid lamellar phase. Simulations of the (31)P NMR spectra were carried out to reveal the formation of different vesicle sizes upon PLB insertion. The bilayer vesicles fragmented into smaller sizes by increasing the concentration of PLB with respect to POPC. Finally, molecular order parameters (S(CD)) were calculated by performing (2)H solid-state NMR studies on deuterated POPC (sn-1 chain) phospholipid bilayers when the PLB peptide was inserted into the membrane.
Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056, USA.
Paresh C
PC
Dave
Elvis K
EK
Tiburu
Krishnan
K
Damodaran
Gary A
GA
Lorigan
D002135:Calcium-Binding Proteins / Q000737:chemistry*; D003903:Deuterium; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000737:chemistry*; D008566:Membranes / Q000737:chemistry*; D008968:Molecular Conformation; D010713:Phosphatidylcholines / Q000737:chemistry*; D010759:Phosphorus Isotopes; D017434:Protein Structure, Tertiary; D000099211:Phospholamban
D002135:Calcium-Binding Proteins; D003903:Deuterium; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010759:Phosphorus Isotopes; D000099211:Phospholamban; C028694:1-palmitoyl-2-oleoylphosphatidylcholine