The interactions of factor V and factor Va light chain with phospholipid vesicles were compared. The results showed that the factor Va light chain bound with the same parameters as factor V when the proteins were present at similar densities on the membrane. The protein-vesicle collisional efficiency was 30-50% for both factor V and factor Va light chain. The factor Va light chain bound at a higher density, and the additional binding interactions had lower affinity. The dissociation process showed negative cooperativity, possibly due to competition for acidic phospholipids in the membrane. The higher molar packing density produced more rapid protein-membrane dissociation rate constants. However, when factor V and Va light chains were present at similar molar densities on the vesicle, the dissociation rates, estimated by two methods, were similar. Analysis of dissociation rates also showed that factor Va interacted with factor Xa on the membrane surface while factor Va light chain did not. Factor Va generated by thrombin digestion of factor V did not result in a major loss of membrane-bound protein mass unless ethylenenediaminetetraacetic acid was present; in the latter case the mass changes indicated that all peptides were removed from the membrane except factor Va light chain. Equilibrium and dynamic measurements showed that ionic strength had a major effect on the dissociation rate but not on the association process. The salt effect indicated interaction between oppositely charged species with the product of the number of charges equal to at least -5.5. Factor Va light chain appeared to interact with phospholipids via a general charge interaction rather than via a specific charge stoichiometry. Lysine modifications readily prevented protein-membrane binding while tryptophan modification had little effect.
|Original language||English (US)|
|Number of pages||9|
|State||Published - 1984|