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Interaction of α-Synuclein and a Cell Penetrating Fusion Peptide with Higher Eukaryotic Cell Membranes Assessed by 19F NMR
journal contribution
posted on 2016-02-21, 16:10 authored by Imola
G. Zigoneanu, Gary J. PielakWe 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.