Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, FL, 32611, USA.
Jiangsu Key Laboratory of New Power Batteries, Jiangsu Collaborative Innovation Center of Biomedical Functional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, 210023, China.
Angew Chem Int Ed Engl. 2017 Sep 18;56(39):11954-11957. doi: 10.1002/anie.201706285. Epub 2017 Aug 25.
Site-selective protein modification is a key step in facilitating protein functionalization and manipulation. To accomplish this, genetically engineered proteins were previously required, but the procedure was laborious, complex, and technically challenging. Herein we report the development of aptamer-based recognition-then-reaction to guide site-selective protein/DNA conjugation in a single step with outstanding selectivity and efficiency. As models, several proteins, including human thrombin, PDGF-BB, Avidin, and His-tagged recombinant protein, were studied, and the results showed excellent selectivity under mild reaction conditions. Taking advantage of aptamers as recognition elements with extraordinary selectivity and affinity, this simple preparation method can tag a protein in a complex milieu. Thus, with the aptamer obtained from cell-SELEX, real-time modification of live-cell membrane proteins can be achieved in one step without any pre-treatment.
位点选择性蛋白质修饰是促进蛋白质功能化和操作的关键步骤。为了实现这一目标，以前需要经过基因工程改造的蛋白质，但该过程繁琐、复杂且技术挑战性大。在此，我们报告了基于适体的识别-反应策略的发展，该策略可在单个步骤中以出色的选择性和效率引导位点选择性蛋白质/DNA 缀合。以几种蛋白质（包括人凝血酶、PDGF-BB、亲和素和 His 标记的重组蛋白）作为模型进行了研究，结果表明在温和的反应条件下具有出色的选择性。利用适体作为具有非凡选择性和亲和力的识别元件，这种简单的制备方法可以在复杂环境中标记蛋白质。因此，利用细胞 SELEX 获得的适体，可以一步实现活细胞膜蛋白质的实时修饰，而无需任何预处理。
