适体
共价键
核酸酶
化学
DNA
电泳剂
生物化学
凝血酶
组合化学
蛋白质检测
靶蛋白
生物物理学
分子生物学
生物
纳米技术
基因
血小板
有机化学
催化作用
材料科学
免疫学
作者
Mary Cacace,Yaniv Tivon,Alexander Deiters
出处
期刊:Methods in Enzymology
日期:2023-01-01
卷期号:: 413-428
标识
DOI:10.1016/bs.mie.2022.08.053
摘要
In this chapter, a new approach to the selective modification of native proteins is discussed, using electrophilic covalent aptamers. These biochemical tools are generated through the site-specific incorporation of a label-transferring or crosslinking electrophile into a DNA aptamer. Covalent aptamers provide the ability to transfer a variety of functional handles to a protein of interest or to irreversibly crosslink to the target. Methods for the aptamer-mediated labeling and crosslinking of thrombin are described. Thrombin labeling is fast and selective, in both simple buffer and in human plasma and outcompetes nuclease-mediated degradation. This approach provides facile, sensitive detection of labeled protein by western blot, SDS-PAGE, and mass spectrometry.
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