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

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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
Volume287
Issue number42
DOIs
Publication statusPublished - 2012 Oct 12
Externally publishedYes

Fingerprint

Lipid bilayers
Lipid Bilayers
Phosphatidylinositols
Nuclear magnetic resonance
Membrane Proteins
Lipids
Proteins
Chemical shift
Membrane Lipids
early endosome antigen 1
Carrier Proteins
Membranes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

NMR analyses of the interaction between the FYVE domain of Early Endosome Antigen 1 (EEA1) and phosphoinositide embedded in a lipid bilayer. / Yokogawa, Mariko; Kobashigawa, Yoshihiro; Yoshida, Naoki; Ogura, Kenji; Harada, Kohsuke; Inagaki, Fuyuhiko.

In: Journal of Biological Chemistry, Vol. 287, No. 42, 12.10.2012, p. 34936-34945.

Research output: Contribution to journalArticle

Yokogawa, Mariko ; Kobashigawa, Yoshihiro ; Yoshida, Naoki ; Ogura, Kenji ; Harada, Kohsuke ; Inagaki, Fuyuhiko. / NMR analyses of the interaction between the FYVE domain of Early Endosome Antigen 1 (EEA1) and phosphoinositide embedded in a lipid bilayer. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 42. pp. 34936-34945.
@article{e9b62b233668404db97349a801675c51,
title = "NMR analyses of the interaction between the FYVE domain of Early Endosome Antigen 1 (EEA1) and phosphoinositide embedded in a lipid bilayer",
abstract = "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.",
author = "Mariko Yokogawa and Yoshihiro Kobashigawa and Naoki Yoshida and Kenji Ogura and Kohsuke Harada and Fuyuhiko Inagaki",
year = "2012",
month = "10",
day = "12",
doi = "10.1074/jbc.M112.398255",
language = "English",
volume = "287",
pages = "34936--34945",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "42",

}

TY - JOUR

T1 - NMR analyses of the interaction between the FYVE domain of Early Endosome Antigen 1 (EEA1) and phosphoinositide embedded in a lipid bilayer

AU - Yokogawa, Mariko

AU - Kobashigawa, Yoshihiro

AU - Yoshida, Naoki

AU - Ogura, Kenji

AU - Harada, Kohsuke

AU - Inagaki, Fuyuhiko

PY - 2012/10/12

Y1 - 2012/10/12

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84867408256&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84867408256&partnerID=8YFLogxK

U2 - 10.1074/jbc.M112.398255

DO - 10.1074/jbc.M112.398255

M3 - Article

C2 - 22915584

AN - SCOPUS:84867408256

VL - 287

SP - 34936

EP - 34945

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 42

ER -