NMR analyses of the interaction between the FYVE domain of Early Endosome Antigen 1 (EEA1) and phosphoinositide embedded in a lipid bilayer

Mariko Yokogawa, Yoshihiro Kobashigawa, Naoki Yoshida, Kenji Ogura, Kohsuke Harada, Fuyuhiko Inagaki

研究成果: Article査読

28 被引用数 (Scopus)

抄録

Phosphoinositides (PIs) are crucial lipid components of membranes and are involved in a number of cellular processes through interactions with their effector proteins. Recently, we have established a lipid-protein nanoscale bilayer (nanodisc) containing PIs, hereafter referred to as PI-nanodisc and demonstrated that it could be used for both qualitative and quantitative evaluations of protein-membrane interactions. Here, we report further NMR analyses for obtaining structural insights at the residue-specific level between PI-binding effector protein and PI-nanodisc, using the FYVE domain of early endosome antigen 1 (EEA1), denoted as EEA1 FYVE, and PI(3)P-nanodisc as a model system. We performed a combination of the NMR analyses including chemical shift perturbation, transferred cross-saturation, and paramagnetic relaxation enhancement experiments. These enabled an identification of the interaction surface, structural change, and relative orientation of EEA1 FYVE to the PI(3)P-incorporated lipid bilayer, substantiating that NMR analyses of protein-membrane interactions using nanodisc makes it possible to show the residue-specific interactions in the lipid bilayer environment.

本文言語English
ページ(範囲)34936-34945
ページ数10
ジャーナルJournal of Biological Chemistry
287
42
DOI
出版ステータスPublished - 2012 10月 12
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 細胞生物学

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