Yahi Nouara, Fantini Jacques
Aix-Marseille Université, PPSN-EA4674, Faculté des Sciences, Marseille, France.
PLoS One. 2014 Aug 20;9(8):e104751. doi: 10.1371/journal.pone.0104751. eCollection 2014.
A broad range of microbial and amyloid proteins interact with cell surface glycolipids which behave as infectivity and/or toxicity cofactors in human pathologies. Here we have deciphered the biochemical code that determines the glycolipid-binding specificity of two major amyloid proteins, Alzheimer's β-amyloid peptide (Aβ) and Parkinson's disease associated protein α-synuclein. We showed that both proteins interact with selected glycolipids through a common loop-shaped motif exhibiting little sequence homology. This 12-residue domain corresponded to fragments 34-45 of α-synuclein and 5-16 of Aβ. By modulating the amino acid sequence of α-synuclein at only two positions in which we introduced a pair of histidine residues found in Aβ, we created a chimeric α-synuclein/Aβ peptide with extended ganglioside-binding properties. This chimeric peptide retained the property of α-synuclein to recognize GM3, and acquired the capacity to recognize GM1 (an Aβ-inherited characteristic). Free histidine (but not tryptophan or asparagine) and Zn2+ (but not Na+) prevented this interaction, confirming the key role of His-13 and His-14 in ganglioside binding. Molecular dynamics studies suggested that the chimeric peptide recognized cholesterol-constrained conformers of GM1, including typical chalice-shaped dimers, that are representative of the condensed cholesterol-ganglioside complexes found in lipid raft domains of the plasma membrane of neural cells. Correspondingly, the peptide had a particular affinity for raft-like membranes containing both GM1 and cholesterol. The chimeric peptide also interacted with several other gangliosides, including major brain gangliosides (GM4, GD1a, GD1b, and GT1b) but not with neutral glycolipids such as GlcCer, LacCer or asialo-GM1. It could inhibit the binding of Aβ1-42 onto neural SH-SY5Y cells and did not induce toxicity in these cells. In conclusion, deciphering the glycolipid code of amyloid proteins allowed us to create a universal ganglioside-binding peptide of only 12-residues with potential therapeutic applications in infectious and neurodegenerative diseases that involve cell surface gangliosides as receptors.
多种微生物蛋白和淀粉样蛋白可与细胞表面糖脂相互作用,这些糖脂在人类疾病中作为感染性和/或毒性辅助因子发挥作用。在此,我们破解了决定两种主要淀粉样蛋白——阿尔茨海默病β-淀粉样肽(Aβ)和帕金森病相关蛋白α-突触核蛋白——糖脂结合特异性的生化密码。我们发现,这两种蛋白均通过一个几乎没有序列同源性的共同环状基序与特定糖脂相互作用。这个由12个残基组成的结构域分别对应于α-突触核蛋白的34 - 45片段和Aβ的5 - 16片段。通过仅在α-突触核蛋白的两个位置调节氨基酸序列,引入Aβ中发现的一对组氨酸残基,我们构建了一种具有扩展神经节苷脂结合特性的嵌合α-突触核蛋白/Aβ肽。这种嵌合肽保留了α-突触核蛋白识别GM3的特性,并获得了识别GM1的能力(这是Aβ遗传的特征)。游离组氨酸(而非色氨酸或天冬酰胺)和Zn2 +(而非Na +)可阻止这种相互作用,证实了His - 13和His - 14在神经节苷脂结合中的关键作用。分子动力学研究表明,嵌合肽识别GM1的胆固醇限制构象,包括典型杯状二聚体,这些构象代表了神经细胞膜脂筏结构域中发现的浓缩胆固醇 - 神经节苷脂复合物。相应地,该肽对同时含有GM1和胆固醇的类脂筏膜具有特殊亲和力。嵌合肽还与其他几种神经节苷脂相互作用,包括主要的脑内神经节苷脂(GM4、GD1a、GD1b和GT1b),但不与中性糖脂如葡萄糖神经酰胺(GlcCer)、乳糖神经酰胺(LacCer)或脱唾液酸GM1相互作用。它可以抑制Aβ1 - 42与神经SH - SY5Y细胞的结合,且在这些细胞中不诱导毒性。总之,破解淀粉样蛋白的糖脂密码使我们能够创建一种仅由12个残基组成的通用神经节苷脂结合肽,在涉及细胞表面神经节苷脂作为受体的感染性和神经退行性疾病中具有潜在治疗应用。