ESR of spin-labeled bacteriophage M13 coat protein in mixed phospholipid bilayers.
Bacteriophage M13 major coat protein was spin-labeled with a nitroxide derivative of iodoacetamide, preferentially at the single methionine that is located in the hydrophobic region of the protein. The spin-labeled protein was incorporated at different lipid-to-protein ratios in phospholipid bilayers composed of dimyristoylphosphatidylglycerol (DMPG), dimyristoylphosphatidylcholine (DMPC), or the 1:1 molar mixture of these lipids. Both conventional and saturation transfer (ST) ESR studies were performed to investigate the rotational motions of the protein over a large dynamic range. The conventional ESR spectra indicate that the mobility of the spin labelled protein in the lipid gel phase decreases in the order: DMPG greater than DMPC/DMPG (1:1) greater than DMPC. In the liquid crystalline phase, the largest mobility is found in DMPC/DMPG (1:1, mol/mol) mixtures, but the mobility is still greater in DMPG than in DMPC. The results are interpreted in terms of different degrees of protein aggregation in the different lipids. Segmental motion with rotational correlation times on the order of tens of nanoseconds, motional anisotropy, and spectral overlap complicate the analysis of the STESR spectra. An estimate of the size of the protein aggregates is found to be in the region of 85 monomer units. Removing the polar tails from the protein by proteolytic digestion results in an enhanced aggregation in the gel phase. In the liquid crystalline phase, the segmental wobbling mobility of the protein is increased relative to the native protein, whereas the overall rotational diffusion is not changed greatly.
de Jongh
H H
HH
Max-Planck-Institut für biophysikalische Chemie, Abteilung Spektroskopie, Göttingen, F.R.G.
Hemminga
M A
MA
Marsh
D
D
D002213:Capsid / Q000648:ultrastructure*; D003090:Coliphages / Q000648:ultrastructure*; D004058:Diffusion; D004134:Dimyristoylphosphatidylcholine; D004578:Electron Spin Resonance Spectroscopy; D007460:Iodoacetamide; D008051:Lipid Bilayers; D009038:Motion; D010715:Phosphatidylglycerols; D010743:Phospholipids; D011485:Protein Binding; D011487:Protein Conformation; D013113:Spin Labels; D013696:Temperature
D008051:Lipid Bilayers; D010715:Phosphatidylglycerols; D010743:Phospholipids; D013113:Spin Labels; C002773:dimyristoylphosphatidylglycerol; D004134:Dimyristoylphosphatidylcholine; D007460:Iodoacetamide