Spin-label ESR of bacteriophage M13 coat protein in mixed lipid bilayers. Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers.
Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of various ratios of dimyristoylphosphatidylcholine (DMPC) to dimyristoylphosphatidylglycerol (DMPG). Spin-label ESR experiments were performed with phospholipids labeled at the C-14 position of the sn-2 chain. For M13 coat protein recombinants with DMPC alone, the relative association constants were determined for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels and found to be 1.0, 1.0, and 2.1 relative to the background DMPC, respectively. The number of association sites for each phospholipid on the protein was found to be 4 per protein monomer. The intrinsic off-rates for lipid exchange at the intramembranous surface of the protein in DMPC alone at 30 degrees C were found to be 5 X 10(6), 6 X 10(6), and 2 X 10(6) s-1 for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels, respectively. Adding DMPG to the DMPC lipid system increased the exchange rates of the lipids on and off the protein. By gel filtration chromatography, it is found that protein aggregation is reduced after addition of DMPG to the lipid system. This is in agreement with measurements of tryptophan fluorescence, which show a decrease in quenching efficiency after introduction of DMPG in the lipid system. The results are interpreted in terms of a model relating the ESR data to the size of the protein-lipid aggregates.
Wolfs
J A
JA
Department of Molecular Physics, Agricultural University, Wageningen, The Netherlands.
Horváth
L I
LI
Marsh
D
D
Watts
A
A
Hemminga
M A
MA
D001435:Bacteriophages / Q000378:metabolism*; D002213:Capsid / Q000378:metabolism*; D002850:Chromatography, Gel; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D005453:Fluorescence; D008051:Lipid Bilayers; D010715:Phosphatidylglycerols / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D013113:Spin Labels; D014364:Tryptophan / Q000378:metabolism
D008051:Lipid Bilayers; D010715:Phosphatidylglycerols; D010743:Phospholipids; D013113:Spin Labels; D014364:Tryptophan; C002773:dimyristoylphosphatidylglycerol; D004134:Dimyristoylphosphatidylcholine