...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biomaterials Science >Immobilization approaches can affect protein dynamics: a surface-enhanced infrared spectroscopic study on lipid-protein interactions
【24h】

Immobilization approaches can affect protein dynamics: a surface-enhanced infrared spectroscopic study on lipid-protein interactions

机译：固定方法可以影响蛋白质动态：表面增强的脂蛋白相互作用的红外光谱研究

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The intrinsically disordered Parkinson disease protein alpha-synuclein (alpha S) performs conformational changes induced by intermolecular protein-protein as well as by protein-membrane interactions. Aggregation of alpha S is a hallmark for the disease, however the role of the membrane in the aggregation process still needs to be clarified. We used a surface-enhanced infrared absorption (SEIRA) spectroscopic approach to investigate the effect of lipid interactions on alpha S conformation. The near-field detection of SEIRA allows to study exclusively structural changes of immobilized alpha S with the advantage that the supernatant remains undetected and thus does not interfere with the spectral read-out. self-assembled monolayer (SAMs) of mixed NHS-PEG-SH linker and MT(PEG)(4) spacer molecules were utilized to immobilize alpha S. The linker/spacer composition of the SAM was adjusted to prevent alpha S-alpha S interactions. Two different methods were applied for site-specific (C-terminal and N-terminal) alpha S immobilization. The immobilized protein was then exposed to lipid vesicles and SEIRA difference spectra were recorded to monitor the alpha S conformation over time. Irrespective of the used immobilization method, alpha S tethering hindered lipid-induced conformational changes. The spectra also indicate that a fraction of the immobilized alpha S eventually desorbs from the surface into the supernatant solution. Desorbed alpha S performs conformational changes and formation of beta-structured aggregates is observed upon interaction with either lipid vesicles or supplementary alpha S. Our study demonstrates that alpha S aggregates only when the protein is free in solution and that surface immobilization procedures, commonly used in many analytical applications, can change the dynamic behavior of proteins thereby affecting protein structure and function.
机译：本质无序的帕金森疾病蛋白α-突触核蛋白（αS）通过分子间蛋白质 - 蛋白和蛋白质 - 膜相互作用进行构象变化。 Alpha S的聚合是疾病的标志，然而，膜在聚集过程中的作用仍然需要澄清。我们使用了表面增强的红外吸收（Seira）光谱方法来研究脂质相互作用对α的构象的影响。 Seira的近场检测允许研究固定的αS的专门结构变化，其优点是上清液未被检测，因此不会干扰光谱读出。利用混合NHS-PEG-SH接头和MT（PEG）（4）间隔物分子的自组装单层（SAMS）固定αS.调节SAM的接头/间隔组合物以防止αS-α的相互作用。施用两种不同的方法，适用于特异性特异性（C末端和N-末端）α的固定化。然后将固定化的蛋白暴露于脂质囊泡，并记录Seira差异光谱以随时间监测αα构象。无论用过的固定方法，alpha s束缚都阻碍了脂质诱导的构象变化。光谱还表明，将固定化αs的一部分最终从表面脱融到上清液溶液中。解吸的αS执行构象变化，并在与脂质囊泡的相互作用时观察到β结构聚集体的形成，我们的研究表明，只有当蛋白质在溶液中没有溶液和该表面固定程序时，常用于α的聚集体许多分析应用，可以改变蛋白质的动态行为，从而影响蛋白质结构和功能。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号