Bane Thomas K, LeBlanc James F, Lee Terry D, Riggs Arthur D
Division of Biology, Beckman Research Institute, The City of Hope National Medical Center, 1450 East Duarte Road, Duarte, CA 91010, USA.
Nucleic Acids Res. 2002 Jul 15;30(14):e69. doi: 10.1093/nar/gnf068.
We report here that surface enhanced laser desorption/ionization-time of flight (SELDI-TOF) mass spectrometry, as performed on a Ciphergen Biosystems ProteinChip System, can be used in conjunction with DNA affinity capture (DACA) to study specific DNA-protein binding. Using DNA molecules bound to a surface, sequence-specific interactions can be detected as demonstrated by a mutation affecting the binding profile for TBP, a transcription factor. Also, a comparison between methylated and unmethylated promoter-containing DNA fragments shows numerous binding profile differences over a mass range extending to >60 kDa. The binding of several proteins is inhibited by methylation of the DNA.
我们在此报告，在Ciphergen生物系统蛋白质芯片系统上进行的表面增强激光解吸/电离飞行时间（SELDI-TOF）质谱分析，可与DNA亲和捕获（DACA）结合使用，以研究特定的DNA-蛋白质结合。使用与表面结合的DNA分子，可以检测到序列特异性相互作用，这通过影响转录因子TBP结合谱的突变得以证明。此外，对含有甲基化和未甲基化启动子的DNA片段的比较显示，在质量范围延伸至>60 kDa的情况下，存在许多结合谱差异。几种蛋白质的结合受到DNA甲基化的抑制。
