Physicochemical characterization of dodecylphosphocholine/palmitoyllysophosphatidic acid/myelin basic protein complexes.
The stoichiometry of dodecylphosphocholine/palmitoyllysophosphatidic acid/myelin basic protein complexes and the location of the protein in the micelles have been investigated by electron paramagnetic resonance, ultracentrifugation, small-angle X-ray scattering, 31P, 13C, and 1H nuclear magnetic resonance spectroscopy, and electron microscopy. Ultracentrifugation measurements indicated that well-defined complexes are formed by association of one protein molecule with approximately 133 detergent molecules. The spin-labels 5-, 12-, and 16-doxylstearate have been incorporated into detergent/protein aggregates. Electron paramagnetic resonance spectral parameters and 13C and 1H nuclear magnetic resonance relaxation times showed that the addition of myelin basic protein does not affect the environment and location of the labels or the organization of the micelles. Previous results suggesting that the protein lies primarily near the surface of the micelles have been confirmed by comparing 13C spectra of the detergents with and without protein with spectra of detergent/protein aggregates containing the spin labels. Electron micrographs of the complexes taken by using the freeze-fracture technique revealed the presence of particles with an estimated radius about three times the radius of the micelles measured by small-angle X-ray scattering. The structural integrity of the complexes appears to be based on intramolecular protein interactions as well as protein-detergent interactions.
Mendz
G L
GL
School of Biochemistry, University of New South Wales, Kensington, Australia.
Miller
D J
DJ
Jamie
I M
IM
White
J W
JW
Brown
L R
LR
Ralston
G B
GB
Kaplin
I J
IJ
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D005614:Freeze Fracturing; D008246:Lysophospholipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008823:Micelles; D008854:Microscopy, Electron / Q000379:methods; D004676:Myelin Basic Protein / Q000737:chemistry*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry; D011817:Rabbits; D013113:Spin Labels; D013229:Stearic Acids; D014461:Ultracentrifugation / Q000379:methods; D014961:X-Ray Diffraction / Q000379:methods
D008246:Lysophospholipids; D008823:Micelles; D004676:Myelin Basic Protein; D013113:Spin Labels; D013229:Stearic Acids; D010767:Phosphorylcholine; C026671:1-palmitoyl-lysophosphatidic acid; C028810:dodecylphosphocholine