Insight into the mechanism of internalization of the cell-penetrating carrier peptide Pep-1 through conformational analysis. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochemistry Année : 2004

Insight into the mechanism of internalization of the cell-penetrating carrier peptide Pep-1 through conformational analysis.

Résumé

Recently, we described a new strategy for the delivery of proteins and peptides into mammalian cells, based on an amphipathic peptide of 21 residues, Pep-1, which was designed on the basis of a protein-interacting domain associated with a nuclear localization sequence and separated by a linker. This peptide carrier constitutes a powerful tool for the delivery of active proteins or peptides both in cultured cells and in vivo, without requiring any covalent coupling. We have examined the conformational states of Pep-1 in its free form and complexed with a cargo peptide and have investigated their ability to interact with phospholipids and the structural consequences of these interactions. From the conformational point of view, Pep-1 behaves significantly differently from other similarly designed cell-penetrating peptides. CD analysis revealed a transition from a nonstructured to a helical conformation upon increase of the concentration. Determination of the structure by NMR showed that in water, its alpha-helical domain extends from residues 4-13. CD and FTIR indicate that Pep-1 adopts a helical conformation in the presence of phospholipids. Adsorption measurements performed at the air-water interface are consistent with the helical form. Pep-1 does not undergo conformational changes upon formation of a particle with a cargo peptide. In contrast, we observe a partial conformational transition when the complex encounters phospholipids. We propose that the membrane crossing process involves formation of a transient transmembrane pore-like structure. Conformational change of Pep-1 is not associated with complexation with its cargo but is induced upon association with the cell membrane.

Dates et versions

hal-00351210 , version 1 (08-01-2009)

Identifiants

Citer

Sébastien Deshayes, Annie Heitz, May C Morris, Pierre Charnet, Gilles Divita, et al.. Insight into the mechanism of internalization of the cell-penetrating carrier peptide Pep-1 through conformational analysis.. Biochemistry, 2004, 43 (6), pp.1449-57. ⟨10.1021/bi035682s⟩. ⟨hal-00351210⟩
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