Molecular Self‐Assembly, Chemical Lithography, and Biochemical Tweezers: A Path for the Fabrication of Functional Nanometer‐Scale Protein Arrays
Molecular Self‐Assembly, Chemical Lithography, and Biochemical Tweezers: A Path for the Fabrication of Functional Nanometer‐Scale Protein Arrays
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分子自组装、化学光刻和生化镊子：功能性纳米级蛋白质阵列的制造途径
DOI：
10.1002/adma.200702189
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发表时间：
2008
影响因子：
29.4
通讯作者：
A. Gölzhäuser
中科院分区：
文献类型：
--
作者：
A. Turchanin;A. Tinazli;M. El;H. Großmann;M. Schnietz;H. Solak;R. Tampé;A. Gölzhäuser
探索约25000个人类基因的无限基因产物和绘制动态蛋白质相互作用网络是当前生物学研究的关键挑战。高度并行的蛋白质芯片技术为这些挑战提供了有前途的工具，甚至可以降低到单分子水平。DNA分子是坚固的，并且很容易以功能方式固定-正如DNA芯片技术的胜利所看到的那样-蛋白质是高度敏感和化学异质性的实体，在体外迅速变性。利用多价性作为设计原则，设计了一个对蛋白质具有极高亲和力的化学识别单元，同时保持了蛋白质的功能。这些多价螯合剂头部由累积的n -硝基三乙酸（NTA）部分组成，通过与过渡金属离子（如Ni(II)）络合，为组氨酸标记的蛋白质提供化学识别。通过添加或去除Ni（II）离子，可以很容易地打开和关闭这些强大的以及特定的结合能力。这些特性使得三- nta螯合剂成为理想的“生化镊子”，用于将蛋白质附着在表面并再次释放它们。3 - ntaunit与6个咪唑基团协调，因此，在纳摩尔范围内的结合亲和力与六组氨酸标签的配位要求完美匹配。为了生成结构蛋白阵列，光刻步骤是必要的。电子束光刻（EBL）和极紫外光干涉光刻（EUV-IL）都是很有前途的技术。EBL以非常可控的方式提供了最大的模式形状和横向尺寸变化。聚焦电子束的应用提供了一个机会，甚至可以产生能够固定单个生物分子的结构。EUV-IL可以在单个处理步骤中在大面积上创建高度并行的图案，其分辨率接近EBL极限，这在纳米技术中具有很大的应用潜力。在本通讯中，我们提出了一种用于蛋白质的横向定义和功能固定的固体模板制造的新途径。该技术是基于电子诱导化学的结合
Exploration of the immensurable gene products of the ∼25000 human genes and mapping of dynamic protein interaction networks are key challenges in current biological research. [1] Highly parallel protein chip technologies deliver promising tools for these challenges even down to the singlemolecule level. [2] Whereas DNA molecules are robust and easy to immobilize in a functional manner—as seen in the triumph of DNA chip technology [3] —proteins are highly sensitive and chemically heterogeneous entities denaturing rapidly in vitro. Using multivalency as a design principle, a chemical recognition unit with exceptionally high affinity for proteins was designed, while maintaining their functionality. [4] These multivalent chelator heads consist of cumulated N-nitrilotriacetic acid (NTA) moieties, which provide chemical recognition for histidine-tagged proteins by complexation of transition metal ions, for example, Ni(II). These strong as well as specific binding capabilities can easily be switched on and off by addition or removal of Ni(II) ions. Such properties make tris-NTA chelators ideal “biochemical tweezers” for attaching proteins to surfaces and releasing them again. A tris-NTAunit coordinates six imidazole moieties and, thus, perfectly matches the coordination demands of a hexahistidine tag with binding affinities in the nanomolar range. [5] For the generation of structured protein arrays, a lithography step is necessary. Both electron-beam lithography (EBL) or extreme UV interference lithography (EUV-IL) are promising for this task. EBL gives the largest variation of pattern shapes and lateral dimensions in a very controlled manner. [6] The application of focused e-beams provides a chance to even generate structures capable of immobilizing single biological molecules. EUV-IL can create highly parallel patterns over large areas in a single processing step with the resolution approaching the EBL limit, [7] which is of great potential for applications in nanotechnology. In this Communication we present a new path for the fabrication of solid templates for the laterally defined and functional immobilization of proteins. The technology is based on the combination of electron-induced chem