Enzyme Activated Gold Nanoparticles for Versatile Site-Selective Bioconjugation
Enzyme Activated Gold Nanoparticles for Versatile Site-Selective Bioconjugation
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DOI：
10.1021/jacs.0c11678
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发表时间：
2021-05-03
影响因子：
15
通讯作者：
Francis, Matthew B.
中科院分区：
文献类型：
--
作者：
Ramsey, Alexandra, V;Bischoff, Amanda J.;Francis, Matthew B.
本文报道了一种新的构建蛋白质和DNA-AuNP偶联物的酶促方法。该策略依赖于aunp与酚类的初始功能化，然后是酪氨酸酶的激活。通过氧化偶联反应，活化的酚与含有脯氨酸、硫醇或苯胺官能团的蛋白质偶联。活化的酚- aunps也与小分子生物素和市售的硫醇- dna结合。这种方法用于AuNP生物偶联的优点包括：(1)初始形成高度稳定的AuNP，可以被酶选择性激活；(2)通过多种功能手柄结合蛋白质或DNA的能力；(3)位点特异性固定；(4)在室温下2小时内完成的简单偶联。将AuNPs的酶氧化偶联应用于烟草花叶病毒(TMV)-AuNP偶联物的构建，并证明了AuNPs与TMV上荧光团之间的能量转移。
A new enzymatic method is reported for constructing protein- and DNA-AuNP conjugates. The strategy relies on the initial functionalization of AuNPs with phenols, followed by activation with the enzyme tyrosinase. Using an oxidative coupling reaction, the activated phenols are coupled to proteins bearing proline, thiol, or aniline functional groups. Activated phenol-AuNPs are also conjugated to a small molecule biotin and commercially available thiol-DNA. Advantages of this approach for AuNP bioconjugation include: (1) initial formation of highly stable AuNPs that can be selectively activated with an enzyme, (2) the ability to conjugate either proteins or DNA through a diverse set of functional handles, (3) site-specific immobilization, and (4) facile conjugation that is complete within 2 h at room temperature under aqueous conditions. The enzymatic oxidative coupling on AuNPs is applied to the construction of tobacco mosaic virus (TMV)-AuNP conjugates, and energy transfer between the AuNPs and fluorophores on TMV is demonstrated.