title: Membrane binding mechanism of an RNA virus-capping enzyme.
abstract: The RNA replication complex of Semliki Forest virus is bound to cytoplasmic membranes via the mRNA-capping enzyme Nsp1. Here we have studied the structure and liposome interactions of a synthetic peptide (245)GSTLYTESRKLLRSWHLPSV(264) corresponding to the membrane binding domain of Nsp1. The peptide interacted with liposomes only if negatively charged lipids were present that induced a structural change in the peptide from a random coil to a partially alpha-helical conformation. NMR structure shows that the alpha-helix is amphipathic, the hydrophobic surface consisting of several leucines, a valine, and a tryptophan moiety (Trp-259). Fluorescence studies revealed that this tryptophan intercalates in the bilayer to the depth of the ninth and tenth carbons of lipid acyl chains. Mutation W259A altered the mode of bilayer association of the peptide and abolished its ability to compete for membrane association of intact Nsp1, demonstrating its crucial role in the membrane association and function of Nsp1.
authors.affiliation: Program in Cellular Biotechnology and NMR Laboratory, Institute of Biotechnology, Viikki Biocenter P. O. Box 56, Helsinki, Finland.
authors.forename: A
authors.initials: A
authors.lastname: Lampio
authors.forename: I
authors.initials: I
authors.lastname: Kilpeläinen
authors.forename: S
authors.initials: S
authors.lastname: Pesonen
authors.forename: K
authors.initials: K
authors.lastname: Karhi
authors.forename: P
authors.initials: P
authors.lastname: Auvinen
authors.forename: P
authors.initials: P
authors.lastname: Somerharju
authors.forename: L
authors.initials: L
authors.lastname: Kääriäinen
mesh_terms: D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000201:enzymology; D008561:Membrane Fusion; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012315:RNA Caps; D012672:Semliki forest virus / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D014364:Tryptophan / Q000737:chemistry
chemical_list: D012315:RNA Caps; D014364:Tryptophan