Kahn Jason S, Xiong Yan, Huang James, Gang Oleg
Center for Functional Nanomaterials, Brookhaven National Laboratory, Upton, New York 11973, United States.
Department of Chemical Engineering, Columbia University, New York, New York 10027, United States.
JACS Au. 2022 Jan 7;2(2):357-366. doi: 10.1021/jacsau.1c00387. eCollection 2022 Feb 28.
DNA nanotechnology has increasingly been used as a platform to scaffold enzymes based on its unmatched ability to structure enzymes in a desired format. The capability to organize enzymes has taken many forms from more traditional 2D pairings on individual scaffolds to recent works introducing enzyme organizations in 3D lattices. As the ability to define nanoscale structure has grown, it is critical to fully deconstruct the impact of enzyme organization at the single-scaffold level. Here, we present an open, three-dimensional (3D) DNA wireframe octahedron which is used to create a library of spatially arranged organizations of glucose oxidase and horseradish peroxidase. We explore the contribution of enzyme spacing, arrangement, and location on the 3D scaffold to cascade activity. The experiments provide insight into enzyme scaffold design, including the insignificance of scaffold sequence makeup on activity, an increase in activity at small enzyme spacings of <10 nm, and activity changes that arise from discontinuities in scaffold architecture. Most notably, the experiments allow us to determine that enzyme colocalization itself on the DNA scaffold dominates over any specific enzyme arrangement.
基于其以所需形式构建酶的无与伦比的能力，DNA纳米技术越来越多地被用作构建酶支架的平台。组织酶的能力有多种形式，从单个支架上更传统的二维配对到最近在三维晶格中引入酶组织的研究。随着定义纳米级结构的能力不断提高，在单支架水平上全面解构酶组织的影响至关重要。在这里，我们展示了一个开放的三维（3D）DNA线框八面体，它被用于创建葡萄糖氧化酶和辣根过氧化物酶空间排列组织的文库。我们探讨了酶间距、排列和在3D支架上的位置对级联活性的贡献。这些实验为酶支架设计提供了见解，包括支架序列组成对活性的无关紧要、在小于10纳米的小酶间距处活性增加以及由支架结构不连续性引起的活性变化。最值得注意的是，这些实验使我们能够确定酶在DNA支架上的共定位本身比任何特定的酶排列更具主导性。
