Department of Chemistry, University College London, 20 Gordon Street, London, WC1H OAJ, UK.
Org Biomol Chem. 2013 Apr 21;11(15):2408-11. doi: 10.1039/c3ob40239h. Epub 2013 Mar 5.
Reversible protein biotinylation is readily affected via conjugation with a bromomaleimide-based reagent followed by reductive cleavage. The intermediate biotinylated protein constructs are stable at physiological temperature and pH 8.0. Quantitative reversibility is elegantly delivered under mild conditions of using a stoichiometric amount of a bis-thiol, thus providing an approach that will be of general interest in chemical biology and proteomics.
通过与溴马来酰亚胺基试剂缀合,然后进行还原裂解,很容易实现蛋白质的可逆生物素化。中间生物素化的蛋白质构建体在生理温度和 pH8.0 下稳定。在使用等摩尔量的双硫醇的温和条件下,实现了定量的可逆性,从而提供了一种在化学生物学和蛋白质组学中普遍感兴趣的方法。