Penketh Philip G, Shyam Krishnamurthy, Baumann Raymond P, Zhu Rui, Ishiguro Kimiko, Sartorelli Alan C, Ratner Elena S
Department of Obstetrics, Gynecology & Reproductive Sciences, Yale University School of Medicine, New Haven, CT 06520-8063, USA.
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520-8066, USA.
Anal Biochem. 2016 Sep 1;508:34-7. doi: 10.1016/j.ab.2016.04.020. Epub 2016 May 14.
Alkylating agents are a significant class of environmental carcinogens as well as commonly used anticancer therapeutics. Traditional alkylating activity assays have utilized the colorimetric reagent 4-(4-nitrobenzyl)pyridine (4NBP). However, 4NBP based assays have a relatively low sensitivity towards harder, more oxophilic alkylating species and are not well suited for the identification of the trapped alkyl moiety due to adduct instability. Herein we describe a method using water as the trapping agent which permits the trapping of simple alkylating electrophiles with a comparatively wide range of softness/hardness and permits the identification of donated simple alkyl moieties.
烷基化剂是一类重要的环境致癌物,也是常用的抗癌治疗药物。传统的烷基化活性测定方法使用比色试剂4-(4-硝基苄基)吡啶(4NBP)。然而,基于4NBP的测定方法对更难、更亲氧的烷基化物种的灵敏度相对较低,并且由于加合物的不稳定性,不太适合鉴定捕获的烷基部分。在此,我们描述了一种使用水作为捕获剂的方法,该方法允许捕获具有较宽软度/硬度范围的简单烷基化亲电试剂,并允许鉴定所提供的简单烷基部分。