Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan.
Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan.
Biochim Biophys Acta Biomembr. 2021 Aug 1;1863(8):183623. doi: 10.1016/j.bbamem.2021.183623. Epub 2021 Apr 30.
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.
神经节苷脂 GM3 位于质膜中，通过阻止表皮生长因子受体（EGFR）的自动磷酸化来抑制细胞生长。生物学研究表明 GM3 与 EGFR 的跨膜结构域相互作用。进一步的生物物理实验对于使用简单的重组系统在双层膜中定量评估肽-糖脂相互作用尤为重要。为了以这种方式检查这些相互作用，我们在 N 端合成了带有硝基苯并二唑荧光团（NBD-TM）的 EGFR 跨膜结构域。基于 NBD-TM 与 ATTO594 标记的 GM3 之间的Förster 共振能量转移（FRET），评估 EGFR 与 GM3 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引起的非特异性相互作用。这种用于选择性检测模型脂质双层中特定脂质-肽相互作用的方法表明，GM3 与 EGFR 跨膜结构域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。
