Calorimetric studies of the structural transitions of the human erythrocyte membrane. Studies of the B and C transitions.
Differential scanning calorimetry has been used to study several structural transitions of the human erythrocyte membrane. Earlier studies have shown that one of these transitions (the A transition) is due to the thermal unfolding of spectrin on the membrane. In this paper, it is shown that two of the other transitions (B and C) exhibit a high sensitivity to a local anesthetic, benzyl alcohol. Increasing the ionic strength of the suspending medium results in a splitting of the B transition into two indepent transitions (B1 and B2). It is found that one of these (B2) is associated with titrating groups, since the midpoint for the transitions shifts by about 20 degrees C, with an apparent pK near 7.5 Extensive bilateral proteolysis by papain causes a drastic decrease in the size of all transitions except the C transition, which remains unaltered. On the other hand, treatment with phospholipase by A2 largely affects the C transition, causing its disappearance. Because of the lack of sensitivity to proteolysis and the high sensitivity to phospholipase, it appears that the C transition has a large extent of 'lipid involvement'. It might result from the melting of a small fraction of phospholipid which exists in a crystalline state under physiological conditions. Alternatively, the C transition could arise from changes in protein-lipid interactions or from lipid-dependent changes in protein-protein interactions, providing one assumes that only protease-resistant portions of membrane proteins are participating.
D002152:Calorimetry, Differential Scanning; D004910:Erythrocyte Membrane / Q000648:ultrastructure*; D004912:Erythrocytes / Q000648:ultrastructure*; D006801:Humans; D009994:Osmolar Concentration; D010206:Papain; D010740:Phospholipases; D011487:Protein Conformation; D013049:Spectrin; D013696:Temperature; D014357:Trypsin
D013049:Spectrin; D010740:Phospholipases; D014357:Trypsin; D010206:Papain