摘要
本文介绍了一种细胞提取液O^6—甲基鸟嘌呤(O^6—MeGua)受体蛋白测定及其底物O^6-[~8H-Me]Gua DNA的制备方法。废物与受体蛋白反应后,甲基从O^6-[~3H-Me]Gua DNA转移到受体蛋白,生成甲基-S-半胱氨酸(Me-S-Cys)。经盐酸水解后,直接测定酸不溶部分的受体蛋白沉淀。方法简便、快速、准确。
An improved method for the quantitation of O6-methylguanine acceptor protein in cell extracts and preparation of ,O69-[3H-methyl] guanine DNA has been described. The methyl group from the O6-methylguanine DNA adduct was transferred to a cysteine residue of acceptor protein after reaction,to yield Me-S-Cys.The protein-bound radioactivity in the acid insoluble fraction was measured following the acid hydrolysis. Data obtained by this assay agree with those obtained by HPLC method, the assay was simple and rapid.
基金
国家自然科学基金
关键词
甲基鸟嘌呤
受体蛋白
氨基酸分析
Acceptor protein O6-[3H-methyl]guanine DNA Amino acid analysis HeLa cell
