A high-sensitivity differential scanning calorimetric study of the interaction of melittin with dipalmitoylphosphatidylcholine fused unilamellar vesicles.
High-sensitivity differential scanning calorimetry has been used to examine the interaction of bee venom melittin with dipalmitoylphosphatidylcholine fused unilamellar vesicles. Experiments were performed under conditions for which melittin in solution is either monomeric (in low salt) or tetrameric (in high salt). It was found that under both sets of conditions melittin abolishes the pretransition at a relatively high lipid-to-protein molar incubation ratio, Ri (about 200) and that at intermediate values of Ri it broadens the main transition profile and reduces the transition enthalpy. This provides evidence which suggests that melittin is at least partially inserted into the apolar region of the bilayer. Evident at low values of Ri are two peaks in the lipid thermal transition profiles, which may arise from a heterogeneous population of lipid vesicles formed through fusion induced by melittin, or by lipid phase separation. For those profiles which exhibited only one peak, transition enthalpies, normalized to those of the lipid in the absence of the protein, are plotted vs. the bound protein-to-lipid molar ratios for the experiments performed under the conditions which give monomeric and tetrameric melittin in solution. These plots yield straight lines, the slopes of which give the number of lipid molecules each protein molecule excludes from participating in the phase transition. These were found to be 9.9 +/- 0.7 and 4.1 +/- 0.5 for monomeric and tetrameric melittin, respectively. The results are discussed in terms of possible models for the binding of melittin to phospholipid vesicles. For simple hexagonal packing of lipid molecules, incorporation as an aggregate is favored when melittin is tetrameric in solution, whereas incorporation as a monomer is favored when melittin is monomeric in solution. For low-salt solutions, evidence is obtained for the contribution of free melittin to lipid fusion, perhaps by the formation of protein bridges between apposed vesicles.
Bradrick
T D
TD
Department of Physics, University of Tennessee, Knoxville 37996-1200.
Freire
E
E
Georghiou
S
S
D015060:1,2-Dipalmitoylphosphatidylcholine; D001514:Bee Venoms / Q000502:physiology*; D002152:Calorimetry, Differential Scanning; D007473:Ion Channels / Q000648:ultrastructure*; D046911:Macromolecular Substances; D008555:Melitten / Q000502:physiology*; D008563:Membrane Lipids; D012492:Salts; D013696:Temperature; D013816:Thermodynamics
D001514:Bee Venoms; D007473:Ion Channels; D046911:Macromolecular Substances; D008563:Membrane Lipids; D012492:Salts; D008555:Melitten; D015060:1,2-Dipalmitoylphosphatidylcholine