Characterization of dodecylphosphocholine/myelin basic protein complexes.
The stoichiometry of myelin basic protein (MBP)/dodecylphosphocholine (DPC) complexes and the location of protein segments in the micelle have been investigated by electron paramagnetic resonance (EPR), ultracentrifugation, photon correlation light scattering, 31P, 13C, and 1H nuclear magnetic resonance (NMR), and electron microscopy. Ultracentrifugation measurements indicate that MBP forms stoichiometrically well-defined complexes consisting of 1 protein molecule and approximately 140 detergent molecules. The spin-labels 5-, 12-, and 16-doxylstearate have been incorporated into DPC/MBP aggregates. EPR spectral parameters and 13C and 1H NMR relaxation times indicate that the addition of MBP does not affect the environment and location of the labels or the organization of the micelles except for a slight increase in size. Previous results indicating that the protein lies primarily near the surface of the micelle have been confirmed by comparing 13C NMR spectra of the detergent with and without protein with spectra of protein/detergent aggregates containing spin-labels. Electron micrographs of the complexes taken by using the freeze-fracture technique confirm the estimated size obtained by light-scattering measurements. Overall, these results indicate that mixtures of MBP and DPC can form highly porous particles with well-defined protein and lipid stoichiometry. The structural integrity of these particles appears to be based on protein-lipid interactions. In addition, electron micrographs of aqueous DPC/MBP suspensions show the formation of a small amount of material consisting of large arrays of detergent micelles, suggesting that MBP is capable of inducing large changes in the overall organization of the detergent.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002794:Choline / Q000031:analogs & derivatives*; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D008854:Microscopy, Electron; D004676:Myelin Basic Protein / Q000378:metabolism*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000378:metabolism; D011485:Protein Binding; D012542:Scattering, Radiation; D013552:Swine; D014461:Ultracentrifugation
D008823:Micelles; D004676:Myelin Basic Protein; D010767:Phosphorylcholine; C028810:dodecylphosphocholine; D002794:Choline