Schweller Ryan M, Constantinou Pamela E, Frankel Nicholas W, Narayan Priyanka, Diehl Michael R
Department of Chemistry, Rice University, 6100 Main Street, MS 142 Houston, Texas 77005, USA.
Bioconjug Chem. 2008 Dec;19(12):2304-7. doi: 10.1021/bc8003606.
A new method for protein surface functionalization was developed that utilizes DNA-conjugated artificial polypeptides to capture recombinant target proteins from the solution phase and direct their deposition onto DNA-functionalized matrices. Protein capture is accomplished through the coiled-coil association of an engineered pair of heterodimeric leucine zippers. Incorporating half of the zipper complex directly into the polypeptides and labeling these polymers with ssDNA enables the polypeptide conjugates to form intermediate linkages that connect the target proteins securely to DNA-functionalized supports. This synthetic route provides an important alternative to conventional DNA-conjugation techniques by allowing proteins to be outfitted site-specifically with ssDNA while minimizing the need for postexpression processing. We demonstrate these attributes by (i) using the capture probes to prepare protein microarrays, (ii) demonstrating control over enzyme activity via deposition of DNA, and, (iii) synthesizing finite-sized, multiprotein complexes that are templated on designed DNA scaffolds in near quantitative yield.
开发了一种蛋白质表面功能化的新方法，该方法利用DNA偶联的人工多肽从溶液相中捕获重组靶蛋白，并将其直接沉积到DNA功能化基质上。蛋白质捕获是通过一对工程化的异源二聚体亮氨酸拉链的卷曲螺旋缔合来完成的。将拉链复合物的一半直接整合到多肽中，并用单链DNA标记这些聚合物，使多肽缀合物能够形成中间连接，将靶蛋白牢固地连接到DNA功能化载体上。这种合成途径为传统的DNA偶联技术提供了一个重要的替代方法，通过允许蛋白质在特定位点配备单链DNA，同时尽量减少表达后处理的需求。我们通过以下方式证明了这些特性：（i）使用捕获探针制备蛋白质微阵列，（ii）通过DNA沉积证明对酶活性的控制，以及（iii）合成以设计的DNA支架为模板的有限大小的多蛋白复合物，产率接近定量。
