title: NMR structure of a viral peptide inserted in artificial membranes: a view on the early steps of the birnavirus entry process.
abstract: Nonenveloped virus must penetrate the cellular membrane to access the cytoplasm without the benefit of membrane fusion. For birnavirus, one of the peptides present in the virus capsid, pep46 for infectious bursal disease virus, is able to induce pores into membranes as an intermediate step of the birnavirus-penetration pathway. Using osmotic protection experiments, we demonstrate here that pep46 and its pore-forming N-terminal moiety (pep22) form pores of different diameters, 5-8 and 2-4 nm, respectively, showing that both pep46 moieties participate to pore formation. The solution structures of pep46, pep22, and pep24 (the pep46 C-terminal moiety) in different hydrophobic environments and micelles determined by (1)H NMR studies provide structural insights of the pep46 domain interaction. In CDCl(3)/CD(3)OH mixture and in dodecylphosphocholine micelles, the N-terminal domain of pep46 is structured in a long kinked helix, although the C terminus is structured in one or two helices depending upon the solvents used. We also show that the folding and the proline isomerization status of pep46 depend on the type of hydrophobic environment. NMR spectroscopy with labeled phospholipid micelles, differential scanning calorimetry, and plasmon waveguide resonance studies show the peptides lie parallel to the lipid-water interface, perturbing the fatty acid chain packing. All these data lead to a model in which the two domains of pep46 interact with the membrane to form pores.
authors.affiliation: Unité de Pharmacologie Chimique et Génétique, CNRS, UMR 8151, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes, 4 Avenue de l'Observatoire, Paris F-75270 Cedex 06, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Galloux
authors.forename: Sonia
authors.initials: S
authors.lastname: Libersou
authors.forename: Isabel D
authors.initials: ID
authors.lastname: Alves
authors.forename: Rodrigue
authors.initials: R
authors.lastname: Marquant
authors.forename: Gilmar F
authors.initials: GF
authors.lastname: Salgado
authors.forename: Human
authors.initials: H
authors.lastname: Rezaei
authors.forename: Jean
authors.initials: J
authors.lastname: Lepault
authors.forename: Bernard
authors.initials: B
authors.lastname: Delmas
authors.forename: Serge
authors.initials: S
authors.lastname: Bouaziz
authors.forename: Nelly
authors.initials: N
authors.lastname: Morellet
mesh_terms: D000818:Animals; D018059:Birnaviridae / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D004912:Erythrocytes / Q000166:cytology / Q000821:virology; D005227:Fatty Acids / Q000737:chemistry; D006461:Hemolysis; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D012756:Sheep; D012997:Solvents / Q000737:chemistry
chemical_list: D005227:Fatty Acids; D008055:Lipids; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides; D011506:Proteins; D012997:Solvents
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