Direct measurement of the energetics of association between myelin basic protein and phosphatidylserine vesicles.
A newly designed high-sensitivity isothermal reaction calorimetry system has been used to investigate the thermodynamics of the association between myelin basic protein and phosphatidylserine vesicles. This instrument has allowed us to measure directly the energetics of the protein-lipid interaction under various conditions. Above the phospholipid phase transition temperature the enthalpy of association is highly exothermic amounting to -160 kcal/mol of protein. Below the phospholipid phase transition temperature the enthalpy of association is exothermic at protein/lipid ratios smaller than 1/50 and endothermic at higher protein/lipid ratios. These studies indicate that the association of myelin basic protein to phosphatidylserine vesicles consists of at least two stages involving different types of binding. The first stage, at low protein/lipid ratios, involves a strong exothermic association of the protein to the membrane and the second, at high protein/lipid ratios, a weaker association probably involving attachment of the protein to the membrane surface only. In the gel phase the second binding stage is endothermic and appears to be correlated with the formation of large vesicle aggregates. This vesicle aggregation is a reversible process dependent upon the physical state of the membrane. The isothermal titration studies have been complemented with high-sensitivity differential scanning calorimetry experiments. It is shown that the dependence of the phospholipid transition enthalpy on the protein/lipid molar ratio can be expressed in terms of the different protein-membrane association enthalpies in the gel and fluid phases of the membrane.
G
G
Ramsay
R
R
Prabhu
E
E
Freire
D000818:Animals; D001921:Brain / Q000378:metabolism; D002151:Calorimetry; D002417:Cattle; D007700:Kinetics; D008081:Liposomes; D004676:Myelin Basic Protein / Q000302:isolation & purification / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism*
D008081:Liposomes; D004676:Myelin Basic Protein; D010718:Phosphatidylserines; C042859:dimyristoylphosphatidylserine