Interaction of melittin with phosphatidylcholine membranes. Binding isotherm and lipid head-group conformation.
The binding of melittin to nonsonicated bilayer membranes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine was studied with an ultracentrifugation assay and with 2H and 31P nuclear magnetic resonance. Melittin binding could best be described by a partition equilibrium with Kp = (2.1 +/- 0.2) X 10(3) M-1, measuring the binding isotherm in the concentration range of 0-100 microM melittin and taking into account electrostatic effects by means of the Gouy-Chapman theory. This partition coefficient is smaller than that deduced for small sonicated vesicles and attests to the tighter lipid packing in the nonsonicated bilayers. Deuterium magnetic resonance revealed a conformational change of the phosphocholine head group upon melittin binding. The quadrupole splittings of the alpha and beta segments of the choline head group varied linearly with the amount of bound melittin but in opposite directions; i.e., the alpha splitting decreased, and the beta splitting increased. This conformational change is not specific to melittin but is a response of the phosphocholine head group to positive membrane surface charges in general. Quantitatively, melittin is one of the most efficient head-group modulators, the efficiency per unit charge comparable to that of charged local anesthetics or hydrophobic ions.
Department of Biophysical Chemistry, Biocenter of the University of Basel, Switzerland.
E
E
Kuchinka
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J
Seelig
D001514:Bee Venoms / Q000378:metabolism*; D001665:Binding Sites; D004563:Electrochemistry; D066298:In Vitro Techniques; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008555:Melitten / Q000378:metabolism*; D008968:Molecular Conformation; D010713:Phosphatidylcholines / Q000378:metabolism*; D013816:Thermodynamics
D001514:Bee Venoms; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D008555:Melitten