title: Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain.
abstract: Although the N terminus of the prion protein (PrP(C)) has been shown to directly associate with lipid membranes, the precise determinants, biophysical basis, and functional implications of such binding, particularly in relation to endogenously occurring fragments, are unresolved. To better understand these issues, we studied a range of synthetic peptides: specifically those equating to the N1 (residues 23-110) and N2 (23-89) fragments derived from constitutive processing of PrP(C) and including those representing arbitrarily defined component domains of the N terminus of mouse prion protein. Utilizing more physiologically relevant large unilamellar vesicles, fluorescence studies at synaptosomal pH (7.4) showed absent binding of all peptides to lipids containing the zwitterionic headgroup phosphatidylcholine and mixtures containing the anionic headgroups phosphatidylglycerol or phosphatidylserine. At pH 5, typical of early endosomes, quartz crystal microbalance with dissipation showed the highest affinity binding occurred with N1 and N2, selective for anionic lipid species. Of particular note, the absence of binding by individual peptides representing component domains underscored the importance of the combination of the octapeptide repeat and the N-terminal polybasic regions for effective membrane interaction. In addition, using quartz crystal microbalance with dissipation and solid-state NMR, we characterized for the first time that both N1 and N2 deeply insert into the lipid bilayer with minimal disruption. Potential functional implications related to cellular stress responses are discussed.
authors.affiliation: Department of Pathology, University of Melbourne, Parkville 3010, Australia.
authors.forename: Martin P
authors.initials: MP
authors.lastname: Boland
authors.forename: Claire R
authors.initials: CR
authors.lastname: Hatty
authors.forename: Frances
authors.initials: F
authors.lastname: Separovic
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Hill
authors.forename: Deborah J
authors.initials: DJ
authors.lastname: Tew
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Barnham
authors.forename: Cathryn L
authors.initials: CL
authors.lastname: Haigh
authors.forename: Michael
authors.initials: M
authors.lastname: James
authors.forename: Colin L
authors.initials: CL
authors.lastname: Masters
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Collins
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000838:Anions / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D000072002:Prion Proteins; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D053835:Unilamellar Liposomes / Q000737:chemistry
chemical_list: D000838:Anions; D008051:Lipid Bilayers; D010455:Peptides; D010743:Phospholipids; D000072002:Prion Proteins; D011328:Prions; C512657:Prnp protein, mouse; D053835:Unilamellar Liposomes