Influence of the lipid composition on the kinetics of concerted insertion and folding of melittin in bilayers.
We have examined the kinetics of the adsorption of melittin, a secondary amphipathic peptide extracted from bee venom, on lipid membranes using three independent and complementary approaches. We probed (i) the change in the polarity of the 19Trp of the peptide upon binding, (ii) the insertion of this residue in the apolar core of the membrane, measuring the 19Trp-fluorescence quenching by bromine atoms attached on lipid acyl chains, and (iii) the folding of the peptide, by circular dichroism (CD). We report a tight coupling of the insertion of the peptide with its folding as an alpha-helix. For all the investigated membrane systems (cholesterol-containing, phosphoglycerol-containing, and pure phosphocholine bilayers), the decrease in the polarity of 19Trp was found to be significantly faster than the increase in the helical content of melittin. Therefore, from a kinetics point of view, the formation of the alpha-helix is a consequence of the insertion of melittin. The rate of melittin folding was found to be influenced by the lipid composition of the bilayer and we propose that this was achieved by the modulation of the kinetics of insertion. The study reports a clear example of the coupling existing between protein penetration and folding, an interconnection that must be considered in the general scheme of membrane protein folding.
Constantinescu
Iren
I
Department of Chemistry, Université de Montréal, CP 6128, Succ. Centre Ville, Montréal, Québec, Canada, H3C 3J7.
Lafleur
Michel
M
D000818:Animals; D001516:Bees; D002784:Cholesterol; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D008555:Melitten / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D014364:Tryptophan
D008051:Lipid Bilayers; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D008555:Melitten; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; D014364:Tryptophan; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine