Site-Selective Protein Immobilization on Polymeric Supports through N-Terminal Imidazolidinone Formation
Site-Selective Protein Immobilization on Polymeric Supports through N-Terminal Imidazolidinone Formation
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DOI：
10.1021/acs.biomac.9b01002
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发表时间：
2019-10-01
影响因子：
6.2
通讯作者：
Francis, Matthew B.
中科院分区：
文献类型：
--
作者：
Koo, Byungjin;Dolan, Nicholas S.;Francis, Matthew B.
蛋白质固定化技术在生物技术和材料科学中有着广泛的应用。以受控的方向连接蛋白质具有固有的优势，但这样做的方法在很大程度上限于半胱氨酸或非规范氨基酸靶向。在此，我们报告了一种方法，通过使用2-吡啶甲醛（2 PCA）衍生物将天然蛋白质的N-末端位置连接到聚合物树脂上。对于高蛋白负载和实际的合成，我们开始这项工作，通过使用Pd催化的交叉偶联胺化制备高反应性的2 PCA衍生物。将所得化合物连接到含胺的聚乙二醇丙烯酰胺树脂（PEGA-NH 2）上，随后与蛋白质的N-末端反应，产生长期稳定的连接，但可以通过加入羟胺逆转。我们设想，这种位点选择性，2 PCA为基础的蛋白质固定化可以提供一个简单的和可推广的固定化协议。
Protein immobilization techniques on polymeric supports have enabled many applications in biotechnology and materials science. Attaching the proteins with controlled orientations has inherent advantages, but approaches for doing this have been largely limited to cysteine or noncanonical amino acid targeting. Herein, we report a method to attach the N-terminal positions of native proteins to polymer resins site-specifically through the use of 2-pyridinecarboxyaldehyde (2PCA) derivatives. For high protein loadings and practical synthesis, we initiated this work by preparing highly reactive 2PCA derivatives using Pd-catalyzed cross-coupling amination. The resulting compounds were attached to amine-containing polyethylene glycol acrylamide resin (PEGA-NH2), which subsequently reacted with the N-termini of proteins to produce linkages that were stable over the long term but could be reversed through the addition of hydroxylamine. We envision that this site-selective, 2PCA-based protein immobilization can provide a simple and generalizable immobilization protocol.