State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, NanKai University, Tianjin 300350, China.
J Mater Chem B. 2021 Jan 7;9(1):80-84. doi: 10.1039/d0tb02302g. Epub 2020 Dec 14.
Specific and expeditious identification and enrichment of target proteins in living cells is often a challenging task. The hexahistidine (6His) tag is frequently used to label artificially engineered proteins produced in prokaryotic or eukaryotic cells. Utilizing the interaction between 6His-tag and nitrilotriacetic acid (NTA) mediated by divalent metal ions (Ni, Cu, Zn or Co), we designed and synthesized a series of Nap-G/Biotin/ANA-FFpYGK-NTA probes that, assisted by alkaline phosphatase (ALP), self-assemble into nanofibers. The probe consists of an NTA group that specifically binds to 6His-tag, an FFpY group that promotes self-assembly facilitated by ALP, and a hydrophobic (Nap-G/ANA/Biotin) capping group for various applications. We demonstrate that the ANA-FFpYGK-NTA(Ni) nanofibers are fit for real-time tracking of His-tagged protein in living cells, and the Biotin-FFpYGK-NTA(Ni) nanofibers are for isolating His-tagged proteins and other proteins that they interact with.
在活细胞中特异性和快速地鉴定和富集靶蛋白通常是一项具有挑战性的任务。六组氨酸（6His）标签常用于标记原核或真核细胞中人工工程化蛋白。利用 6His 标签与二价金属离子（Ni、Cu、Zn 或 Co）介导的亚氨基二乙酸（NTA）之间的相互作用，我们设计并合成了一系列 Nap-G/生物素/ANA-FFpYGK-NTA 探针，这些探针在碱性磷酸酶（ALP）的辅助下自组装成纳米纤维。该探针由一个特异性结合 6His 标签的 NTA 基团、一个由 ALP 促进自组装的 FFpY 基团和一个疏水性（Nap-G/ANA/生物素）封端基团组成，可用于各种应用。我们证明，ANA-FFpYGK-NTA（Ni）纳米纤维适合实时跟踪活细胞中 His 标记蛋白，而 Biotin-FFpYGK-NTA（Ni）纳米纤维适合分离 His 标记蛋白及其相互作用的其他蛋白。
