FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers

Mikito Nakano; Shinya Hanashima; Toshiaki Hara; Kazuya Kabayama; Yuya Asahina; Hironobu Hojo; Naoko Komura; Hiromune Ando; Thomas K.M. Nyholm; J. Peter Slotte; Michio Murata

doi:10.1016/j.bbamem.2021.183623

FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers

Mikito Nakano
, Shinya Hanashima^*
, Toshiaki Hara
, Kazuya Kabayama
, Yuya Asahina
, Hironobu Hojo
, Naoko Komura
, Hiromune Ando
, Thomas K.M. Nyholm
, J. Peter Slotte
, Michio Murata

^*Tämän työn vastaava kirjoittaja

Tutkimustuotos: Lehtiartikkeli › Artikkeli › Tieteellinen › vertaisarvioitu

16 Sitaatiot (Scopus)

Abstrakti

Ganglioside GM3 in the plasma membranes suppresses cell growth by preventing the autophosphorylation of the epidermal growth factor receptor (EGFR). Biological studies have suggested that GM3 interacts with the transmembrane segment of EGFR. Further biophysical experiments are particularly important for quantitative evaluation of the peptide-glycolipid interplay in bilayer membranes using a simple reconstituted system. To examine these interactions in this way, we synthesized the transmembrane segment of EGFR bearing a nitrobenzoxadiazole fluorophore (NBD-TM) at the N-terminus. The affinity between EGFR and GM3 was evaluated based on Förster resonance energy transfer (FRET) between NBD-TM and ATTO594-labeled GM3 in bilayers where their non-specific interaction due to lateral proximity was subtracted by using NBD-labeled phospholipid. This method for selectively detecting the specific lipid-peptide interactions in model lipid bilayers disclosed that the lateral interaction between GM3 and the transmembrane segment of EGFR plays a certain role in disturbing the formation of active EGFR dimers.

Alkuperäiskieli	Englanti
Artikkeli	183623
Julkaisu	Biochimica et Biophysica Acta - Biomembranes
Vuosikerta	1863
Numero	8
DOI - pysyväislinkit	https://doi.org/10.1016/j.bbamem.2021.183623
Tila	Julkaistu - 1 elok. 2021
OKM-julkaisutyyppi	A1 Julkaistu artikkeli, soviteltu

Rahoitus

This work was partially supported by KAKENHI Grant Numbers, JP16H06315 (M. M.) JP17H06406 (M. M.) and JP19K05713 (S. H.) from Japan Society for the Promotion of Science (JSPS) and CREST Grant Number JPMJCR18H2 (S.H. and H.A.) from Japan Science and Technology Agency (JST). We thank Prof. Kenichi GN Suzuki (Gifu University) for helpful discussions. M.N. thanks Tobitate! (Leap for Tomorrow) Young Ambassador Program from MEXT, Japan for a financial support to study abroad in the Slotte group.

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10.1016/j.bbamem.2021.183623

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Nakano, M., Hanashima, S., Hara, T., Kabayama, K., Asahina, Y., Hojo, H., Komura, N., Ando, H., Nyholm, T. K. M., Slotte, J. P., & Murata, M. (2021). FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers. Biochimica et Biophysica Acta - Biomembranes, 1863(8), Artikkeli 183623. https://doi.org/10.1016/j.bbamem.2021.183623

@article{a35e30dea54d443bb665c758ac1a78bc,

title = "FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers",

abstract = "Ganglioside GM3 in the plasma membranes suppresses cell growth by preventing the autophosphorylation of the epidermal growth factor receptor (EGFR). Biological studies have suggested that GM3 interacts with the transmembrane segment of EGFR. Further biophysical experiments are particularly important for quantitative evaluation of the peptide-glycolipid interplay in bilayer membranes using a simple reconstituted system. To examine these interactions in this way, we synthesized the transmembrane segment of EGFR bearing a nitrobenzoxadiazole fluorophore (NBD-TM) at the N-terminus. The affinity between EGFR and GM3 was evaluated based on F{\"o}rster resonance energy transfer (FRET) between NBD-TM and ATTO594-labeled GM3 in bilayers where their non-specific interaction due to lateral proximity was subtracted by using NBD-labeled phospholipid. This method for selectively detecting the specific lipid-peptide interactions in model lipid bilayers disclosed that the lateral interaction between GM3 and the transmembrane segment of EGFR plays a certain role in disturbing the formation of active EGFR dimers.",

keywords = "EGF receptor, FRET, Ganglioside GM3, Lipid-peptide interaction, Transmembrane helix",

author = "Mikito Nakano and Shinya Hanashima and Toshiaki Hara and Kazuya Kabayama and Yuya Asahina and Hironobu Hojo and Naoko Komura and Hiromune Ando and Nyholm, \{Thomas K.M.\} and Slotte, \{J. Peter\} and Michio Murata",

note = "Funding Information: This work was partially supported by KAKENHI Grant Numbers, JP16H06315 (M. M.) JP17H06406 (M. M.) and JP19K05713 (S. H.) from Japan Society for the Promotion of Science (JSPS) and CREST Grant Number JPMJCR18H2 (S.H. and H.A.) from Japan Science and Technology Agency (JST). Funding Information: We thank Prof. Kenichi GN Suzuki (Gifu University) for helpful discussions. M.N. thanks Tobitate! (Leap for Tomorrow) Young Ambassador Program from MEXT, Japan for a financial support to study abroad in the Slotte group. Publisher Copyright: {\textcopyright} 2021",

year = "2021",

month = aug,

day = "1",

doi = "10.1016/j.bbamem.2021.183623",

language = "English",

volume = "1863",

journal = "Biochimica et Biophysica Acta - Biomembranes",

issn = "0005-2736",

publisher = "Elsevier",

number = "8",

}

Nakano, M, Hanashima, S, Hara, T, Kabayama, K, Asahina, Y, Hojo, H, Komura, N, Ando, H, Nyholm, TKM, Slotte, JP & Murata, M 2021, 'FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers', Biochimica et Biophysica Acta - Biomembranes, Vuosikerta 1863, Nro 8, 183623. https://doi.org/10.1016/j.bbamem.2021.183623

TY - JOUR

T1 - FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers

AU - Nakano, Mikito

AU - Hanashima, Shinya

AU - Hara, Toshiaki

AU - Kabayama, Kazuya

AU - Asahina, Yuya

AU - Hojo, Hironobu

AU - Komura, Naoko

AU - Ando, Hiromune

AU - Nyholm, Thomas K.M.

AU - Slotte, J. Peter

AU - Murata, Michio

N1 - Funding Information: This work was partially supported by KAKENHI Grant Numbers, JP16H06315 (M. M.) JP17H06406 (M. M.) and JP19K05713 (S. H.) from Japan Society for the Promotion of Science (JSPS) and CREST Grant Number JPMJCR18H2 (S.H. and H.A.) from Japan Science and Technology Agency (JST). Funding Information: We thank Prof. Kenichi GN Suzuki (Gifu University) for helpful discussions. M.N. thanks Tobitate! (Leap for Tomorrow) Young Ambassador Program from MEXT, Japan for a financial support to study abroad in the Slotte group. Publisher Copyright: © 2021

PY - 2021/8/1

Y1 - 2021/8/1

N2 - Ganglioside GM3 in the plasma membranes suppresses cell growth by preventing the autophosphorylation of the epidermal growth factor receptor (EGFR). Biological studies have suggested that GM3 interacts with the transmembrane segment of EGFR. Further biophysical experiments are particularly important for quantitative evaluation of the peptide-glycolipid interplay in bilayer membranes using a simple reconstituted system. To examine these interactions in this way, we synthesized the transmembrane segment of EGFR bearing a nitrobenzoxadiazole fluorophore (NBD-TM) at the N-terminus. The affinity between EGFR and GM3 was evaluated based on Förster resonance energy transfer (FRET) between NBD-TM and ATTO594-labeled GM3 in bilayers where their non-specific interaction due to lateral proximity was subtracted by using NBD-labeled phospholipid. This method for selectively detecting the specific lipid-peptide interactions in model lipid bilayers disclosed that the lateral interaction between GM3 and the transmembrane segment of EGFR plays a certain role in disturbing the formation of active EGFR dimers.

AB - Ganglioside GM3 in the plasma membranes suppresses cell growth by preventing the autophosphorylation of the epidermal growth factor receptor (EGFR). Biological studies have suggested that GM3 interacts with the transmembrane segment of EGFR. Further biophysical experiments are particularly important for quantitative evaluation of the peptide-glycolipid interplay in bilayer membranes using a simple reconstituted system. To examine these interactions in this way, we synthesized the transmembrane segment of EGFR bearing a nitrobenzoxadiazole fluorophore (NBD-TM) at the N-terminus. The affinity between EGFR and GM3 was evaluated based on Förster resonance energy transfer (FRET) between NBD-TM and ATTO594-labeled GM3 in bilayers where their non-specific interaction due to lateral proximity was subtracted by using NBD-labeled phospholipid. This method for selectively detecting the specific lipid-peptide interactions in model lipid bilayers disclosed that the lateral interaction between GM3 and the transmembrane segment of EGFR plays a certain role in disturbing the formation of active EGFR dimers.

KW - EGF receptor

KW - FRET

KW - Ganglioside GM3

KW - Lipid-peptide interaction

KW - Transmembrane helix

UR - https://www.scopus.com/pages/publications/85105314858

U2 - 10.1016/j.bbamem.2021.183623

DO - 10.1016/j.bbamem.2021.183623

M3 - Article

C2 - 33933428

AN - SCOPUS:85105314858

SN - 0005-2736

VL - 1863

JO - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

IS - 8

M1 - 183623

ER -

FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers

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