Negative cooperativity and aggregation in biphasic binding of mastoparan X peptide to membranes with acidic lipids.

The change of Trp fluorescence intensity when large vesicles with 10% acidic lipid are added to mastoparan X solutions reflects a fast and a slow binding process. By means of a novel procedure of data analysis that takes advantage of so-called mass conservation plots we have separated association isotherms related to: (i) the apparent fast pre-equilibrium; and (ii) the final equilibrium, respectively. This approach also reveals that the intrinsic fluorescence signal of the slow binding is considerably raised against that of the fast binding, presumably indicating a penetration of bound peptide from the lipid/water interface into the apolar lipid core. The shape of either binding curve discloses a pronounced tendency of aggregation. Furthermore, it turns out that in the slow process the final binding ratio decreases markedly compared with the initial fast binding ratio. Accordingly the occupation of final binding sites must exert a substantial effect of negative cooperativity on the affinity of the interfacial binding states.