一种快速灵敏的测定苯并[a]芘与蛋白质共价结合的方法。

A rapid and sensitive method for determination of covalent binding of benzo[a]pyrene to proteins.

作者信息

Wallin H, Schelin C, Tunek A, Jergil B

出版信息

Chem Biol Interact. 1981 Dec;38(1):109-18. doi: 10.1016/0009-2797(81)90157-5.

Abstract

A method is presented for the quantitative determination of covalent binding of metabolically activated benzo[a]pyrene to microsomal proteins. After incubation of radiolabelled benzo[a]pyrene with microsomes and NADPH, the mixture is applied to filter paper discs. These are immersed in ethanol to precipitate the proteins. Unbound radiolabel is removed by repeated washes of the filters in organic solvents before scintillation counting. The method is simple, rapid, sensitive and accurate, and works both with 14C- and 3H-labelled compounds. The method is suitable for measuring the incorporation of other radiolabelled xenobiotics to proteins of both microsomes and other subcellular fractions and for the analysis of binding to isolated proteins.

摘要

本文介绍了一种定量测定代谢活化的苯并[a]芘与微粒体蛋白共价结合的方法。将放射性标记的苯并[a]芘与微粒体及NADPH一起孵育后，将混合物点样到滤纸圆片上。将这些滤纸圆片浸入乙醇中使蛋白质沉淀。在进行闪烁计数之前，通过在有机溶剂中反复洗涤滤纸去除未结合的放射性标记。该方法简单、快速、灵敏且准确，适用于14C和3H标记的化合物。该方法适用于测量其他放射性标记的外源化合物掺入微粒体和其他亚细胞组分的蛋白质中，以及分析与分离蛋白的结合情况。