Influence of the lipid composition on the kinetics of concerted insertion and folding of melittin in bilayers |
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Authors: | Iren Constantinescu |
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Institution: | Department of Chemistry, Université de Montréal, C.P. 6128, Succ. Centre Ville, Montréal, Québec, Canada, H3C 3J7 |
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Abstract: | 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 α-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 α-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. |
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Keywords: | CD circular dichroism chol cholesterol POPC 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine POPG 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol 6 7Br2PC 1-(6 7-dibromo)palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine LUV large unilamellar vesicles PC phosphocholine Ri incubation lipid/melittin molar ratio SUV small unilamellar vesicles PS phosphatidylserine DMPC dimyristoyl-sn-glycero-3-phosphocholine DPPC dipalmitoyl-sn-glycero-3-phosphocholine DOPC dioleoyl-sn-glycero-3-phosphocholine DOPG dioleoyl-sn-glycero-3-phosphoglycerol |
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