Fukui Kenji, Takahata Yoshio, Nakagawa Noriko, Kuramitsu Seiki, Masui Ryoji
RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan.
Nucleic Acids Res. 2007;35(15):e100. doi: 10.1093/nar/gkm575. Epub 2007 Aug 7.
Electrospray ionization with Fourier-transform ion cyclotron resonance mass spectrometry (ESI-FT ICR MS) is a powerful tool for analyzing the precise structural features of biopolymers, including oligonucleotides. Here, we described the detailed characterization of a newly discovered nuclease activity of the C-terminal domain of Thermus thermophilus MutS2 (ttMutS2). Using this method, the length, nucleotide content and nature of the 5'- and 3'-termini of the product oligonucleotides were accurately identified. It is revealed that the C-terminal domain of ttMutS2 incised the phosphate backbone of oligodeoxynucleotides non-sequence-specifically at the 3' side of the phosphates. The simultaneous identification of the innumerable fragments was achieved by the extremely high-accuracy of ESI-FT ICR MS.
傅里叶变换离子回旋共振质谱联用的电喷雾电离（ESI-FT ICR MS）是分析包括寡核苷酸在内的生物聚合物精确结构特征的有力工具。在此，我们描述了嗜热栖热菌MutS2（ttMutS2）C端结构域新发现的核酸酶活性的详细特征。使用该方法，可准确鉴定产物寡核苷酸5'和3'末端的长度、核苷酸含量及性质。结果表明，ttMutS2的C端结构域在磷酸基团的3'侧非序列特异性地切割寡脱氧核苷酸的磷酸主链。通过ESI-FT ICR MS的极高准确性实现了对无数片段的同时鉴定。
