Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain

Martin Boland, Claire R Hatty, Frances Separovic, Andrew F Hill, Deborah J Tew, Kevin Jeffrey Barnham, Cathryn Louise Haigh, Michael James, Colin L Masters, Steven John Collins

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Although the N terminus of the prion protein (PrPC) 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 PrPC 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.
Original languageEnglish
Pages (from-to)32282-32292
Number of pages11
JournalJournal of Biological Chemistry
Volume285
Issue number42
DOIs
Publication statusPublished - 15 Oct 2010
Externally publishedYes

Fingerprint

Quartz Crystal Microbalance Techniques
Phospholipids
Quartz crystal microbalances
Peptides
PrPC Proteins
Lipids
Unilamellar Liposomes
Phosphatidylglycerols
Lipid bilayers
Terminal Repeat Sequences
Endosomes
Phosphatidylserines
Lipid Bilayers
Membrane Lipids
Phosphatidylcholines
Fluorescence
Nuclear magnetic resonance
Membranes
Processing
Prions

Cite this

Boland, Martin ; Hatty, Claire R ; Separovic, Frances ; Hill, Andrew F ; Tew, Deborah J ; Barnham, Kevin Jeffrey ; Haigh, Cathryn Louise ; James, Michael ; Masters, Colin L ; Collins, Steven John. / Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 42. pp. 32282-32292.
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abstract = "Although the N terminus of the prion protein (PrPC) 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 PrPC 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.",
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Boland, M, Hatty, CR, Separovic, F, Hill, AF, Tew, DJ, Barnham, KJ, Haigh, CL, James, M, Masters, CL & Collins, SJ 2010, 'Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain', Journal of Biological Chemistry, vol. 285, no. 42, pp. 32282-32292. https://doi.org/10.1074/jbc.M110.123398

Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain. / Boland, Martin; Hatty, Claire R; Separovic, Frances; Hill, Andrew F; Tew, Deborah J; Barnham, Kevin Jeffrey; Haigh, Cathryn Louise; James, Michael; Masters, Colin L; Collins, Steven John.

In: Journal of Biological Chemistry, Vol. 285, No. 42, 15.10.2010, p. 32282-32292.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Anionic phospholipid interactions of the prion protein N terminus are minimally perturbing and not driven solely by the octapeptide repeat domain

AU - Boland, Martin

AU - Hatty, Claire R

AU - Separovic, Frances

AU - Hill, Andrew F

AU - Tew, Deborah J

AU - Barnham, Kevin Jeffrey

AU - Haigh, Cathryn Louise

AU - James, Michael

AU - Masters, Colin L

AU - Collins, Steven John

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