Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao-Tong University School of Medicine , Shanghai 200025, China.
J Chem Inf Model. 2013 Sep 23;53(9):2360-8. doi: 10.1021/ci4002487. Epub 2013 Aug 21.
The small ubiquitin-related modifier (SUMO)-specific protease (SENP) processes SUMOs to mature forms and deconjugates them from various modified substrates. Loss of the equilibrium from desumoylation catalyzed by abnormal SENP1 is associated with cancers and transcription factor activity. In spite of the significant role of SENP1, the molecular basis of its desumoylation remains unclear. Here, MD simulations and QM/MM methods are combined to investigate the catalytic mechanism of desumoylation. The results showed that substrate SUMO1-RanGAP1 fitted into the catalytic pocket of SENP1 by the break of internal hydrophobic interactions and the isomerization of isopeptide from trans to cis. After that, the nucleophilic sulfur anion of Cys603 in SENP1 attacked the carbonyl carbon of Gly97 of SUMO1 to trigger the reaction, and then a tetrahedral intermediate and an acyl-enzyme intermediate were generated in turn, leading to the final release of enzyme SENP1 and two products, free SUMO1 and RanGAP1. In the process, nucleophilic attack was identified as the rate-determining step with a potential energy barrier of 20.2 kcal/mol. These results are in agreement with experimental data from mutagenesis and other experiments. Our findings elucidate the catalytic mechanism of SENP1 with its substrate and may provide a better understanding of SENP desumoylation. In particular, we have identified key residues in SENP1 needed for desumoylation that might be beneficial for the design of novel inhibitors of SENP1-related diseases.
小泛素相关修饰物(SUMO)特异性蛋白酶(SENP)将 SUMO 加工成熟形式,并从各种修饰底物中去除它们。异常 SENP1 催化的去 SUMOylation 失去平衡与癌症和转录因子活性有关。尽管 SENP1 具有重要作用,但去 SUMOylation 的分子基础仍不清楚。在这里,我们结合 MD 模拟和 QM/MM 方法研究了去 SUMOylation 的催化机制。结果表明,底物 SUMO1-RanGAP1 通过内部疏水相互作用的破坏和异肽从反式到顺式的异构化,适合 SENP1 的催化口袋。之后,SENP1 中的 Cys603 的亲核硫阴离子攻击 SUMO1 的 Gly97 的羰基碳,引发反应,然后依次生成四面体中间物和酰基-酶中间物,最终导致 SENP1 和两种产物,游离 SUMO1 和 RanGAP1 的释放。在这个过程中,亲核攻击被确定为速率决定步骤,其势能垒为 20.2 kcal/mol。这些结果与突变和其他实验的实验数据一致。我们的发现阐明了 SENP1 与其底物的催化机制,可能为 SENP 去 SUMOylation 提供了更好的理解。特别是,我们已经确定了 SENP1 中用于去 SUMOylation 的关键残基,这可能有助于设计新型 SENP1 相关疾病的抑制剂。