論文

査読有り 国際誌
2013年5月14日

FRET-based imaging of transbilayer movement of pepducin in living cells by novel intracellular bioreductively activatable fluorescent probes.

Organic & biomolecular chemistry
  • Mieko Tsuji
  • ,
  • Satoshi Ueda
  • ,
  • Tasuku Hirayama
  • ,
  • Kensuke Okuda
  • ,
  • Yoshiaki Sakaguchi
  • ,
  • Aoi Isono
  • ,
  • Hideko Nagasawa

11
18
開始ページ
3030
終了ページ
7
記述言語
英語
掲載種別
DOI
10.1039/c3ob27445d

To elucidate the mechanisms of direct transmembrane penetration of pepducins, which are artificial lipopeptide G protein-coupled receptor (GPCR) modulators, we developed two types of FRET-based probes, Pep13-FL-SS-Dab (13) targeting the inner leaflet of the lipid bilayer and Pep13-Dab-SS-FL (14) targeting the cytosol, respectively. They are composed of a pepducin moiety and a fluorescent switch component consisting of 5(6)-carboxyfluorescein (FAM) as a fluorophore and dabcyl as a quencher connected through disulfide bond linkage. When they are internalized into the cytosol, intracellular glutathione can cleave the disulfide bond to release the quencher, which results in a turn-on fluorescence signal. Using these probes, we performed live cell imaging of transbilayer movements of pepducins on MCF-7 cells for the first time. The results suggested that the lipid moiety of the probes facilitated pepducin flipping across and tethering to the membrane. The present study raises the possibility of applying the probe architecture for direct intracellular drug delivery.

リンク情報
DOI
https://doi.org/10.1039/c3ob27445d
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/23532512
ID情報
  • DOI : 10.1039/c3ob27445d
  • PubMed ID : 23532512

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