Functional immobilization of the small GTPase Rab6A on DNA-gold nanoparticles by using a site-specifically attached poly(ethylene glycol) linker and thiol place-exchange reaction
Functional immobilization of the small GTPase Rab6A on DNA-gold nanoparticles by using a site-specifically attached poly(ethylene glycol) linker and thiol place-exchange reaction
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DOI：
10.1002/cbic.200600422
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发表时间：
2007-01-02
期刊：
影响因子：
3.2
通讯作者：
Niemeyer, Christof M.
中科院分区：
文献类型：
--
作者：
Becker, Christian F. W.;Marsac, Yoann;Niemeyer, Christof M.
近年来，在无机纳米颗粒上实现蛋白质和 DNA 等生物大分子功能性固定的有效策略已成为人们日益关注的主题，因为它们构成了功能纳米材料合理设计的基础。 [1, 2] 这些材料在生物分析和生物分子电子学中具有广泛的应用范围。 [3-5] 然而，蛋白质在纳米颗粒上的功能性固定通常很难实现，因为蛋白质中存在多个官能团使连接任务变得复杂。它们附着在表面上，同时保持其功能齐全。[6]常用的方法通常利用非特异性吸收或共价附着。[7, 8]这些策略通常会导致纳米颗粒表面上蛋白质的不受控制的固定和随机方向。克服这些方法产生的潜在问题的一个策略是基于蛋白质和功能化纳米颗粒表面之间的特异性相互作用。[9, 10]我们在这里报告了共价连接的蛋白质-纳米颗粒缀合物的受控组装，通过一种基于用功能化聚乙二醇连接分子[11]通过天然化学连接反应对蛋白质进行高度特异性修饰的方法（方案1）。[12]这种基于 PEG 的连接体具有化学惰性和水溶性；[13] 这使得它们作为暴露于溶剂的间隔物非常有吸引力，可以通过附加的硫醇基团轻松与纳米颗粒表面发生反应。在本研究中，我们使用 Rab 家族的小型 GTP 酶研究了这一策略。 Rab 蛋白是中心
Efficient strategies to achieve functional immobilization of biomacromolecules, such as proteins and DNA, on inorganic nanoparticles have been the subject of growing interest in recent years, since they form a basis for the rational design of functional nanomaterials.[1, 2] These materials have a broad scope of application in bioanalytics and biomolecular electronics.[3–5] However, the functional immobilization of proteins on nanoparticles is often difficult to achieve because the occurrence of multiple functional groups in proteins complicates the task of linking them to surfaces while keeping them fully functional.[6] Commonly employed methods often take advantage of nonspecific absorption or covalent attachment.[7, 8] These strategies often lead to uncontrolled immobilization and random orientation of the proteins on the nanoparticle surface. One strategy to overcome the potential problems arising from these approaches is based on specific interactions between the protein and a functionalized nanoparticle surface.[9, 10]We report here on the controlled assembly of covalently linked protein–nanoparticle conjugates through an approach based on the highly specific modification of proteins with a functionalized poly (ethylene glycol) linker molecule [11] by a native chemical ligation reaction (Scheme1).[12] Such PEG-based linkers are chemically inert and water soluble;[13] this renders them highly attractive as solvent-exposed spacers that can easily react with nanoparticle surfaces through an appended thiol group. In this study, we investigated this strategy by using a small GTPase of the Rab family. Rab proteins are cen-