FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers
FRET detects lateral interaction between transmembrane domain of EGF receptor and ganglioside GM3 in lipid bilayers
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FRET 检测 EGF 受体跨膜结构域与脂质双层中神经节苷脂 GM3 之间的横向相互作用
DOI：
10.1016/j.bbamem.2021.183623
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Murata Michio
中科院分区：
文献类型：
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作者：
Nakano Mikito;Hanashima Shinya;Hara Toshiaki;Kabayama Kazuya;Asahina Yuya;Hojo Hironobu;Komura Naoko;Ando Hiromune;Nyholm Thomas K.M.;Slotte J. Peter;Murata Michio
质膜中的神经节苷脂GM 3通过阻止表皮生长因子受体（EGFR）的自磷酸化来抑制细胞生长。生物学研究表明，GM 3与EGFR的跨膜区段相互作用。进一步的生物物理实验是特别重要的定量评价的肽-糖脂的相互作用，在双层膜使用一个简单的重建系统。为了以这种方式检查这些相互作用，我们合成了在N-末端带有硝基苯并恶二唑荧光团（NBD-TM）的EGFR的跨膜区段。基于双层中NBD-TM和ATTO 594标记的GM 3之间的Förster共振能量转移（FRET）评价EGFR和GM 3之间的亲和力，其中通过使用NBD标记的磷脂减去由于侧向接近而引起的非特异性相互作用。这种选择性检测模型脂质双层中特异性脂-肽相互作用的方法揭示了GM 3与EGFR跨膜片段之间的侧向相互作用在干扰活性EGFR二聚体的形成中起一定作用。
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.