通过基质辅助激光解吸电离飞行时间质谱法分析铑嵌入剂的位点特异性DNA光裂解

Site-specific DNA photocleavage by rhodium intercalators analyzed by MALDI-TOF mass spectrometry.

作者信息

Brunner Jens, Barton Jacqueline K

机构信息

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.

出版信息

J Am Chem Soc. 2006 May 31;128(21):6772-3. doi: 10.1021/ja0612753.

DOI:10.1021/ja0612753

PMID:16719441

Abstract

The DNA photocleavage reaction of mismatch-selective Rh complexes has been analyzed by MALDI-TOF mass spectrometry as well as gel electrophoresis analysis of radioactively tagged oligonucleotides. Analogous results are obtained with these two techniques showing site-specific cleavage neighboring the mismatch to yield primarily 5'- and 3'-phosphate termini. Additional intermediates and products are observed, however, using MALDI-TOF analysis. MALDI-TOF mass spectrometry is seen to be particularly powerful in the analysis of DNA cleavage by site-specific molecules. The method requires no radioactive labeling, only little material, and analysis can be accomplished within minutes. Moreover, this mass spectral analysis of DNA cleavage yields direct information regarding products rather than simply the base pair site of cleavage.

摘要

通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）以及对放射性标记寡核苷酸的凝胶电泳分析，对错配选择性铑配合物的DNA光裂解反应进行了分析。这两种技术得到了类似的结果，表明在错配附近发生位点特异性裂解，主要产生5'-和3'-磷酸末端。然而，使用MALDI-TOF分析观察到了额外的中间体和产物。MALDI-TOF质谱在分析位点特异性分子对DNA的裂解方面特别有效。该方法不需要放射性标记，所需材料很少，并且可以在几分钟内完成分析。此外，这种对DNA裂解的质谱分析可直接提供有关产物的信息，而不仅仅是裂解的碱基对位点信息。