Department of Biochemistry, Chemistry Institute, São Paulo University, São Paulo, SP, Brazil.
PLoS One. 2013 Jul 22;8(7):e67194. doi: 10.1371/journal.pone.0067194. Print 2013.
Alzheimer's disease (AD) is characterized by brain accumulation of the neurotoxic amyloid-β peptide (Aβ) and by loss of cholinergic neurons and nicotinic acetylcholine receptors (nAChRs). Recent evidence indicates that memory loss and cognitive decline in AD correlate better with the amount of soluble Aβ than with the extent of amyloid plaque deposits in affected brains. Inhibition of nAChRs by soluble Aβ40 is suggested to contribute to early cholinergic dysfunction in AD. Using phage display screening, we have previously identified a heptapeptide, termed IQ, homologous to most nAChR subtypes, binding with nanomolar affinity to soluble Aβ40 and blocking Aβ-induced inhibition of carbamylcholine-induced currents in PC12 cells expressing α7 nAChRs. Using alanine scanning mutagenesis and whole-cell current recording, we have now defined the amino acids in IQ essential for reversal of Aβ40 inhibition of carbamylcholine-induced responses in PC12 cells, mediated by α7 subtypes and other endogenously expressed nAChRs. We further investigated the effects of soluble Aβ, IQ and analogues of IQ on α3β4 nAChRs recombinantly expressed in HEK293 cells. Results show that nanomolar concentrations of soluble Aβ40 potently inhibit the function of α3β4 nAChRs, and that subsequent addition of IQ or its analogues does not reverse this effect. However, co-application of IQ makes the inhibition of α3β4 nAChRs by Aβ40 reversible. These findings indicate that Aβ40 inhibits different subtypes of nAChRs by interacting with specific receptor domains homologous to the IQ peptide, suggesting that IQ may be a lead for novel drugs to block the inhibition of cholinergic function in AD.
阿尔茨海默病(AD)的特征是大脑中神经毒性淀粉样β肽(Aβ)的积累,以及胆碱能神经元和烟碱型乙酰胆碱受体(nAChRs)的丧失。最近的证据表明,AD 中的记忆丧失和认知能力下降与可溶性 Aβ的量相关性更好,而与受影响大脑中淀粉样斑块沉积的程度相关性较差。可溶性 Aβ40 对 nAChRs 的抑制被认为有助于 AD 早期的胆碱能功能障碍。通过噬菌体展示筛选,我们之前已经鉴定出一种七肽,称为 IQ,与大多数 nAChR 亚型同源,与可溶性 Aβ40 以纳摩尔亲和力结合,并阻断 Aβ诱导的 PC12 细胞中α7 nAChRs 表达的烟碱诱导电流的抑制。通过丙氨酸扫描突变和全细胞电流记录,我们现在已经确定了 IQ 中对逆转可溶性 Aβ40 抑制 PC12 细胞中烟碱诱导反应所必需的氨基酸,该反应由α7 亚型和其他内源性表达的 nAChRs 介导。我们进一步研究了可溶性 Aβ、IQ 和 IQ 类似物对在 HEK293 细胞中重组表达的α3β4 nAChRs 的影响。结果表明,纳摩尔浓度的可溶性 Aβ40 可强烈抑制α3β4 nAChRs 的功能,而随后添加 IQ 或其类似物并不能逆转这种效应。然而,IQ 的共同应用使 Aβ40 对α3β4 nAChRs 的抑制作用变得可逆。这些发现表明,Aβ40 通过与 IQ 肽同源的特定受体结构域相互作用抑制不同亚型的 nAChRs,这表明 IQ 可能是一种新型药物的先导,用于阻断 AD 中胆碱能功能的抑制。