Chemical Synthesis, Crystal Structure and Enzymatic Evaluation of a Dinucleotide Spore Photoproduct Analogue Containing a Formacetal Linker
Chemical Synthesis, Crystal Structure and Enzymatic Evaluation of a Dinucleotide Spore Photoproduct Analogue Containing a Formacetal Linker
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DOI：
10.1002/chem.201101821
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发表时间：
2011-08-01
影响因子：
4.3
通讯作者：
Li, Lei
中科院分区：
文献类型：
--
作者：
Lin, Gengjie;Chen, Chun-Hsing;Li, Lei
孢子光产物（SP）是细菌内生孢子中唯一的DNA光损伤产物。它的光形成和修复由金属酶孢子光产物裂解酶（SPL）组成的独特的SP生物化学。尽管SP的发现已有近50年的历史，但其晶体结构仍不清楚，结构信息的缺乏极大地阻碍了SP生物化学的研究。利用缩甲醛连接物和有机合成方法，我们成功地制备了一个二核苷酸SP电子等排体5 R-CH（2）SP，它在两个胸腺嘧啶残基之间含有一个中性的CH（2）部分，而不是一个磷酸。中性连接体极大地促进了结晶过程，使我们能够在发现半个世纪后获得这种有趣的胸腺嘧啶二聚体的晶体结构。对这种5 R-CH（2）SP化合物的进一步ROESY光谱、DFT计算和酶研究证明它具有与5 R-SP物质相似的性质，这表明所揭示的结构真实地反映了自然界中产生的SP的结构。
Spore photoproduct (SP) is the exclusive DNA photodamage product found in bacterial endospores. Its photoformation and repair by a metalloenzyme spore photoproduct lyase (SPL) composes the unique SP biochemistry. Despite the fact that the SP was discovered almost 50 years ago, its crystal structure is still unknown and the lack of structural information greatly hinders the study of SP biochemistry. Employing a formacetal linker and organic synthesis, we successfully prepared a dinucleotide SP isostere 5R-CH(2)SP, which contains a neutral CH(2) moiety between the two thymine residues instead of a phosphate. The neutral linker dramatically facilitates the crystallization process, allowing us to obtain the crystal structure for this intriguing thymine dimer half a century after its discovery. Further ROESY spectroscopic, DFT computational, and enzymatic studies of this 5R-CH(2)SP compound prove that it possesses similar properties with the 5R-SP species, suggesting that the revealed structure truly reflects that of SP generated in Nature.