Peptide models of the helical hydrophobic transmembrane segments of membrane proteins: interactions of acetyl-K2-(LA)12-K2-amide with phosphatidylethanolamine bilayer membranes.

High-sensitivity differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy were used to study the interaction of a synthetic alpha-helical hydrophobic transmembrane peptide, acetyl-Lys(2)-(Leu-Ala)(12)-Lys(2)-amide [(LA)(12)], and members of a homologous series of n-saturated diacylphosphatidylethanolamines (PEs). In the lower range of peptide mole fractions, the DSC endotherms exhibited by the lipid/peptide mixtures consist of two components. The temperature and cooperativity of the sharper, higher temperature component are very similar to those of pure PE bilayers and are almost unaffected by variations in the protein/lipid ratio. However, the fractional contribution of this component to the total enthalpy changes decreases with increases in peptide concentration, and this component completely disappears at higher protein mole fractions. The other component, which is less cooperative and occurs at a lower temperature, predominates at higher protein concentrations. These two components of the DSC endotherm have been assigned to the chain-melting phase transitions of peptide-nonassociated and peptide-associated PE molecules, respectively. Although the temperature at which the peptide-associated PE molecules melt is progressively decreased by increases in (LA)(12) concentration, the magnitude of this downward shift is progressively greater as the length of the PE hydrocarbon chain decreases. As well, mixtures of (LA)(12) with the longer chain PEs exhibit unusual biomodal enthalpy variations, suggesting peptide immiscibility in thicker gel state bilayers. Moreover, the enthalpy of the chain-melting transition of the peptide-associated PE does not decrease to zero even at high peptide concentrations, indicating that (LA)(12) attenuates but does not abolish the cooperative gel/liquid-crystalline phase transition of the lipids with which it is in contact. Our FTIR spectroscopic data indicate that (LA)(12) remains in a predominantly alpha-helical conformation in liquid-crystalline PE bilayers of various hydrophobic thickness but that the helical conformation is altered in gel-state PE bilayers generally, probably due to peptide lateral aggregation. These data also suggest that (LA)(12) significantly disorders the hydrocarbon chains of adjacent PE molecules in both the gel and liquid-crystalline states, relatively independently of lipid hydrocarbon chain length. Many aspects of PE/(LA)(12) interactions exhibit a different dependence on the hydrophobic thickness of the host bilayer than was observed in our previous study of (LA)(12)-phosphatidylcholine (PC) model membranes [Zhang et al. (1995) Biochemistry 34, 2362-2371]. The differing effects of (LA)(12) incorporation on PE and PC bilayers is ascribed primarily to the much stronger lipid polar headgroup interactions characteristic of the former system. Finally, the considerable differences observed in the behavior of (LA)(12) and the related polyleucine-based peptide P(24) in both PC and PE bilayers indicate that the structure of the hydrophobic core of alpha-helical transmembrane peptides can affect their conformational plasticity and state of aggregation and thus the nature of their interactions with different phospholipid bilayers.