Saridaki Theodora, Zampagni Mariagioia, Mannini Benedetta, Evangelisti Elisa, Taddei Niccolò, Cecchi Cristina, Chiti Fabrizio
Department of Biochemical Sciences, University of Florence, Viale Morgagni 50, 50134 Florence, Italy.
J Mol Biol. 2012 Aug 24;421(4-5):616-30. doi: 10.1016/j.jmb.2012.02.007. Epub 2012 Feb 9.
A group of diverse human pathologies is associated with proteins unable to retain their native state and convert into prefibrillar and fibrillar amyloid aggregates that are then deposited in the extracellular space. Glycosaminoglycans (GAGs) have been found to physically associate with these deposits and also to promote their formation in vitro. However, the effect of GAGs on the toxicity of these aggregates has been investigated in only one protein system, the amyloid β peptide associated with Alzheimer's disease. In this study, we investigate whether GAGs affect the toxicity of the N-terminal domain of Escherichia coli HypF (HypF-N) oligomers on Chinese hamster ovarian cells and the mechanism by which such suppression is mediated. The results show that heparin and other GAGs inhibit the toxicity observed by HypF-N oligomers in a dose-dependent manner. GAGs were not found to bind preformed HypF-N oligomers, change their morphological and structural characteristics or disaggregate them. Nevertheless, they were found to bind to the cells' surface and prevent the interaction of the oligomers with the cells. Overall, the results indicate that GAGs have a generic ability to inhibit the toxicity of aberrant protein oligomers and that such toxicity suppression can occur through different mechanisms, such as through binding to the oligomers with consequent loss of interaction of the oligomers to the GAGs present on the cell surface, as proposed previously for amyloid β aggregates, or through mechanisms independent of direct GAG-oligomer binding, as shown here for HypF-N aggregates.
一组多样的人类病理学与无法保持其天然状态并转化为原纤维和纤维状淀粉样聚集体的蛋白质有关,这些聚集体随后沉积在细胞外空间。已发现糖胺聚糖(GAGs)与这些沉积物发生物理结合,并在体外促进其形成。然而,仅在一种蛋白质系统(即与阿尔茨海默病相关的淀粉样β肽)中研究了GAGs对这些聚集体毒性的影响。在本研究中,我们调查了GAGs是否影响大肠杆菌HypF(HypF-N)寡聚体的N端结构域对中国仓鼠卵巢细胞的毒性以及这种抑制作用的介导机制。结果表明,肝素和其他GAGs以剂量依赖的方式抑制HypF-N寡聚体所观察到的毒性。未发现GAGs与预先形成的HypF-N寡聚体结合、改变其形态和结构特征或使其解聚。然而,发现它们与细胞表面结合并阻止寡聚体与细胞的相互作用。总体而言,结果表明GAGs具有抑制异常蛋白质寡聚体毒性的一般能力,并且这种毒性抑制可以通过不同机制发生,例如通过与寡聚体结合,从而导致寡聚体与细胞表面存在的GAGs的相互作用丧失,如先前对淀粉样β聚集体所提出的,或通过独立于直接GAG-寡聚体结合的机制,如此处对HypF-N聚集体所示。