title: Interaction of α-synuclein and a cell penetrating fusion peptide with higher eukaryotic cell membranes assessed by ¹⁹F NMR.
abstract: We show that fluorine NMR can be used to monitor the insertion and change in conformation of a ¹⁹F-labeled cell-penetrating peptide upon interacting with the cellular plasma membrane. α-Synuclein and a construct comprising a cell-penetrating peptide covalently attached to its N-terminus were studied. Important information about the interaction of the proteins with CHO-K1 cells was obtained by monitoring the diminution of ¹⁹F resonances of 3-fluoro-l-tyrosine labeled proteins. For α-synuclein, a decrease in the resonance from position 39 was observed indicating that only the N-terminal third region of the protein interacts with plasma membrane. However, when the fusion construct was incubated with the cells, a decrease in the resonance from the fusion peptide region was noted with no change in the resonances from α-synuclein region. Longer incubation, studied by using confocal fluorescence microscopy, revealed that the fusion construct translocates into the cells, but α-synuclein alone did not cross the membrane in significant amounts.
authors.affiliation: Department of Chemistry, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Imola G
authors.initials: IG
authors.lastname: Zigoneanu
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Pielak
mesh_terms: D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D006224:Cricetinae; D005057:Eukaryotic Cells / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011485:Protein Binding; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics
chemical_list: D057846:Cell-Penetrating Peptides; D051844:alpha-Synuclein