文摘
A high-performance affinity purification technique has been developed for cisplatin (CDDP)-damagedDNA binding proteins directly from crude nuclear extracts of HeLaS3 cell using novel submicronbeads synthesized by copolymerization of styrene and glycidyl methacrylate (GMA). The beadsdramatically decreased both nonspecific protein adsorption on solid surfaces and elution volume andsimplified the handling procedure. Preparation of the beads for purification was carried out byimmobilization of telomeric repeats, (TTAGGG)n, on the surface after the reaction with CDDP. Atleast nine proteins clearly showed higher affinity to CDDP-DNA and were identified by amino acidsequence analysis including HMGB (high mobility group), hUBF (human upstream binding factor),and Ku autoantigen, which were previously reported to be components of CDDP-damaged DNA bindingproteins.