Solubilization of myelin membranes by detergents.
The major proteins of myelin have classically been extracted in organic solvents. Here we investigated some of the characteristics of brain myelin solubilization in aqueous detergent solutions. At comparable molar concentrations, two nonionic detergents, i.e., octyl glucoside and Lubrol PX, proved relatively better myelin solubilizers than the detergents related to the bile salts, i.e., cholate and CHAPS. The two former detergents solubilized more protein than lipid and the two latter ones more lipid than protein from myelin membranes. All four detergents solubilized the phospholipid more efficiently than the cholesterol component of myelin. The detergent concentrations required for myelin solubilization were reduced substantially if the temperature and the salt concentration of the media were increased. As much as 3 mg of lyophilized myelin (about 1 mg of protein) were solubilized readily per milliliter of a solution containing 30 mM octyl glucoside and 0.1 M sodium sulfate in 0.1 M sodium phosphate buffer, pH 6.7. Each of the detergents studied, including the above four, sodium dodecyl sulfate (SDS). Triton X-100, and Zwittergent 3-14, had its own advantages and drawbacks as myelin protein extractors. The nonionic amphiphiles and CHAPS left a small residue mainly composed of proteins of the Wolfgram fraction, as revealed by SDS-polyacrylamide gel electrophoresis. Octyl glucoside was preferred, given its versatility as solubilizer, ultraviolet transparency, and high critical micellar concentration. Observations on possible difficulties that may be encountered are also included.
Aveldaño
M I
MI
Instituto Santiago Ramón y Cajal, C.S.I.C., Madrid, Spain.
Díaz
R S
RS
Regueiro
P
P
Monreal
J
J
D000818:Animals; D002417:Cattle; D002498:Centrifugation; D002784:Cholesterol / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D008566:Membranes / Q000378:metabolism; D009185:Myelin Proteins / Q000378:metabolism; D018991:Myelin Proteolipid Protein; D009186:Myelin Sheath / Q000378:metabolism*; D009994:Osmolar Concentration; D010743:Phospholipids / Q000378:metabolism; D012995:Solubility; D013696:Temperature
D003902:Detergents; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D010743:Phospholipids; D002784:Cholesterol