DNA聚合酶η小分子抑制剂筛选  

Screening of small molecule inhibitors targeting DNA polymeraseη

作　　者：曹佳佳 叶舒迈 赵烨 CAO Jiajia;YE Shumai;ZHAO Ye(College of Life Sciences,Zhejiang University,Hangzhou 310058,Zhejiang,China)

机构地区：[1]浙江大学生命科学学院,浙江杭州310058

出　　处：《浙江大学学报（农业与生命科学版）》2024年第1期35-41,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：浙江省自然科学基金项目(LDQ23C050002)。

摘　　要：DNA损伤修复以及基因组稳定性维持对于动植物正常生长和防御逆境至关重要。针对DNA聚合酶错误掺入导致的基因组不稳定性,本研究以DNA聚合酶η为研究对象,通过计算分子模拟对接的方式,对其可能的小分子抑制剂进行筛选,并对其酶动力学参数进行测定。结果显示:脱氧腺苷三磷酸(deoxyadenosine triphosphate,d ATP)对DNA聚合酶η的活性具有抑制效果,使其延伸的相对效率为36%~42%。分子模拟对接和体外实验结果表明,相较于dATP(亲和力为-26.7 kJ/mol),环鸟苷酸-腺苷酸(cyclic GMP-AMP,cGAMP)与DNA聚合酶η具有更低的结合能(亲和力为-35.1 kJ/mol)。酶动力学参数测定结果也表明,相较于dATP,cGAMP具有更强的抑制能力且在浓度为0.5 mmol/L时达到最强(相对延伸效率为13%)。因此,本研究筛选获得了针对DNA聚合酶η的一种潜在的小分子抑制剂。同时,鉴于该蛋白质高表达导致细胞对抗肿瘤药物(DNA损伤剂)的耐受性,这为新型药物的开发提供了依据。The repair of DNA damage and maintenance of genomic stability are essential for the normal growth and adverse defense of plants and animals.In view of the genomic instability caused by the misincorporation of DNA polymerase,this study took DNA polymeraseηas the research object and screened its possible small molecule inhibitors by computational simulated molecular docking and detected their enzyme kinetic parameters.The results showed that deoxyadenosine triphosphate(dATP)had an inhibitory effect on the activity of DNA polymeraseη,resulting in a relative extension efficiency of 36%to 42%.Simulated molecular docking and in vitro experimental results showed that cyclic GMP-AMP(cGAMP)had a lower binding energy(with an affinity of-35.1 kJ/mol)than dATP(with an affinity of-26.7 kJ/mol)to DNA polymeraseη.Enzyme kinetic experiments also showed that cGAMP had a stronger inhibitory ability than dATP and achieved the maximum effect at the concentration of 0.5 mmol/L(with a relative extension efficiency of 13%).Therefore,a potential small molecule inhibitor targeting DNA polymeraseηwas screened out in this study.At the same time,in view of the tolerance to antitumor drug(DNA damage agent)caused by high expression of this protein,these results provide a basis for the development of new drugs.

关 键 词：DNA损伤修复 DNA聚合酶 酶动力学 计算生物学 环鸟苷酸-腺苷酸 

分 类 号：Q513[生物学—生物化学]