Structural parameters of the myelin transmembrane proteolipid in reverse micelles.
The Folch-Pi proteolipid is the most abundant structural protein from the central nervous system myelin. This protein-lipid complex, normally insoluble in water, requires only a small amount of water for solubilization in reverse micelles of sodium bis (2-ethylhexyl) sulfosuccinate (AOT) in isooctane. The characterization of the proteolipid-free and proteolipid-containing micelles was undertaken by light scattering and fluorescence recovery after fringe pattern photobleaching (FRAPP) experiments. Quasi elastic light scattering (QELS) was carried out at a high (200 mM) AOT concentration, at low water-to-surfactant mole ratio (Wo = 7) and at increasing protein occupancy. Two apparent hydrodynamic radii, differing tenfold in size, were obtained from correlation functions. The smaller one (RaH = 5.2 nm) remains constant and corresponds to that measured for protein-free micelles. The larger one increases linearly with protein concentration. In contrast, FRAPP measurements of self-diffusion coefficients were found unaffected by the proteolipid concentration. Accordingly, they have been performed at constant protein/surfactant mole ratios. The equivalent RH, extrapolated to zero AOT concentration for protein-free reverse micelles (2.9 nm) and in the presence of the proteolipid (4.6 nm), do not reveal the mode of organization previously suggested by QELS measurements. The complex picture emerging from this work represents a first step in the characterization of an integral membrane protein in reverse micelles.
Binks
B P
BP
Laboratoire de Spectroscopie Hertzienne, Paris, France.
Chatenay
D
D
Nicot
C
C
Urbach
W
W
Waks
M
M
D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008027:Light; D008823:Micelles; D008961:Models, Structural; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D010777:Photochemistry; D011487:Protein Conformation; D012542:Scattering, Radiation; D013861:Thiocyanates
D005452:Fluoresceins; D005456:Fluorescent Dyes; D008823:Micelles; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate