Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University, Boston, MA 02115, USA.
J Mass Spectrom. 2012 Apr;47(4):490-501. doi: 10.1002/jms.2054.
The analysis of DNA adducts is of importance in understanding DNA damage, and in the last few years mass spectrometry (MS) has emerged as the most comprehensive and versatile tool for routine characterization of modified oligonucleotides. The structural analysis of modified oligonucleotides, although routinely analyzed using mass spectrometry, is followed by a large amount of data, and a significant challenge is to locate the exact position of the adduct by computational spectral interpretation, which still is a bottleneck. In this report, we present an additional feature of the in-house developed GenoMass software, which determines the exact location of an adduct in modified oligonucleotides by connecting tandem mass spectrometry (MS/MS) to a combinatorial isomer library generated in silico for nucleic acids. The performance of this MS/MS approach using GenoMass software was evaluated by MS/MS data interpretation for an unadducted and its corresponding N-acetylaminofluorene (AAF) adducted 17-mer (5'OH-CCT ACC CCT TCC TTG TA-3'OH) oligonucleotide. Further computational screening of this AAF adducted 17-mer oligonucleotide (5'OH-CCT ACC CCT TCC TTG TA-3'OH) from a complex oligonucleotide mixture was performed using GenoMass. Finally, GenoMass was also used to identify the positional isomers of the AAF adducted 15-mer oligonucleotide (5'OH-ATGAACCGGAGGCCC-3'OH). GenoMass is a simple, fast, data interpretation software that uses an in silico constructed library to relate the MS/MS sequencing approach to identify the exact location of adduct on oligonucleotides.
DNA 加合物分析对于理解 DNA 损伤至关重要,近年来,质谱(MS)已成为常规鉴定修饰寡核苷酸的最全面和通用的工具。尽管使用质谱法对修饰寡核苷酸进行了结构分析,但随后会产生大量数据,因此一个重大挑战是通过计算谱解释来确定加合物的确切位置,这仍然是一个瓶颈。在本报告中,我们介绍了内部开发的 GenoMass 软件的另一个功能,该功能通过将串联质谱(MS/MS)与组合异构体文库连接,确定修饰寡核苷酸中加合物的确切位置,该文库是在计算机上为核酸生成的。通过对未加合物及其相应的 N-乙酰氨基芴(AAF)加合物 17 mer(5'OH-CCT ACC CCT TCC TTG TA-3'OH)寡核苷酸的 MS/MS 数据解释来评估此 MS/MS 方法使用 GenoMass 的性能。使用 GenoMass 对复杂寡核苷酸混合物中的这种 AAF 加合物 17mer 寡核苷酸(5'OH-CCT ACC CCT TCC TTG TA-3'OH)进行了进一步的计算筛选。最后,还使用 GenoMass 来鉴定 AAF 加合物 15mer 寡核苷酸(5'OH-ATGAACCGGAGGCCC-3'OH)的位置异构体。GenoMass 是一种简单、快速的数据分析软件,它使用计算机构建的文库将 MS/MS 测序方法联系起来,以确定寡核苷酸上加合物的确切位置。