Different states of self-association of melittin in phospholipid bilayers. A resonance energy transfer approach.
Melittin is known to self-associate as tetramers in solutions of high ionic strength. Here, an N-bromosuccinimide oxidized-Trp19 melittin is prepared. This derivative can act as an acceptor of the fluorescence of native melittin and is used in order to observe a possible self-association of melittin in phospholipid bilayers. Resonance energy transfer was shown to occur in solutions of high ionic strength, showing that oxidized melittin can associate with native melittin. In phospholipid bilayers, no association is detected in the absence of NaCl. In its presence, an equilibrium between monomeric melittin and oligomeric species is observed. These species are not dimers, but any other degree of association may account for our experimental results. Significant differences in characteristic transfer efficiency reveal differences in the structure of these oligomers according to the length or state of phospholipids (fluid or at the transition temperature). These bound complexes are also different from the soluble hetero-oligomer. Some models of bound complexes are proposed which may explain the leakage and the further disruption of vesicles or cells induced by melittin.
Talbot
J C
JC
Centre de Recherche Paul Pascal, C.N.R.S., Domaine Universitaire, Talence, France.
Faucon
J F
JF
Dufourcq
J
J
D015060:1,2-Dipalmitoylphosphatidylcholine; D001514:Bee Venoms; D001974:Bromosuccinimide; D004735:Energy Transfer; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008555:Melitten; D008958:Models, Molecular; D008968:Molecular Conformation; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D001514:Bee Venoms; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D008555:Melitten; D015060:1,2-Dipalmitoylphosphatidylcholine; D001974:Bromosuccinimide