Membrane insertion and dissociation processes of a model transmembrane helix.
An important subject for elucidating membrane protein (MP) folding is how transmembrane helices (TMHs) insert into and dissociate from membranes. We investigated helix dissociation kinetics and insertion topology by means of intervesicular transfer of the fluorophore-labeled completely hydrophobic model transmembrane helix NBD-(LALAAAA)(3)-NH(2) (NBD = 7-nitro-2-1,3-benzoxadiazol-4-yl). The peptide forms a topologically stable transmembrane helix, which is in a monomer-antiparallel dimer equilibrium [Yano, Y., Takemoto, T., Kobayashi, S., Yasui, H., Sakurai, H., Ohashi, W., Niwa, M., Futaki, S., Sugiura, Y., and Matsuzaki, K. (2002) Biochemistry 41, 3073-3080]. The helix transfer kinetics, representing the helix dissociation process, was monitored by fluorescence recovery of the quenched peptide in donor vesicles containing a quencher upon its transfer to acceptor vesicles without the quencher. The transfer kinetics and vesicle concentration dependence demonstrated that the transfer was mediated by monomer in the aqueous phase. Furthermore, the activation enthalpy was estimated to be +17.7 +/- 1.3 kcal mol(-1). Helix insertion topology, detected by chemical quenching of the NBD group in the outer leaflet by dithionite ions, was found to be controlled by transmembrane electric potential-helix macro dipole interaction. On the basis of these observations, a model for the helix insertion/dissociation processes was discussed.
D000409:Alanine / Q000737:chemistry; D019277:Entropy; D031541:Fluorescence Resonance Energy Transfer; D007930:Leucine / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008564:Membrane Potentials; D008567:Membranes, Artificial; D008956:Models, Chemical; D008958:Models, Molecular; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D017433:Protein Structure, Secondary; D021381:Protein Transport; D013696:Temperature
D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010455:Peptides; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine; D007930:Leucine; D000409:Alanine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine