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

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24 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)34936-34945
Number of pages10
JournalJournal of Biological Chemistry
Issue number42
Publication statusPublished - 2012 Oct 12


ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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