Interaction of 伪-Synuclein and a Cell Penetrating Fusion Peptide with Higher Eukaryotic Cell Membranes Assessed by 19F NMR
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- 作者：Imola G. Zigoneanu ; Gary J. Pielak
- 刊名：Molecular Pharmaceutics
- 出版年：2012
- 出版时间：April 2, 2012
- 年：2012
- 卷：9
- 期：4
- 页码：1024-1029
- 全文大小：271K
- 年卷期：v.9,no.4(April 2, 2012)
- ISSN：1543-8392
文摘
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. 伪-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 伪-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 伪-synuclein region. Longer incubation, studied by using confocal fluorescence microscopy, revealed that the fusion construct translocates into the cells, but 伪-synuclein alone did not cross the membrane in significant amounts.