Exploring the Potential of Arginine to Increase Coelenterazine-Renilla Luciferase Affinity and Enzyme Stability: Kinetic and Molecular Dynamics Studies
{"title":"Exploring the Potential of Arginine to Increase Coelenterazine-Renilla Luciferase Affinity and Enzyme Stability: Kinetic and Molecular Dynamics Studies","authors":"Maryam Salehian, Rahman Emamzadeh, Mahboobeh Nazari","doi":"10.1007/s10930-024-10208-x","DOIUrl":null,"url":null,"abstract":"<div><p><i>Renilla</i> luciferase catalyzes the oxidation of coelenterazine to coelenteramide and results in the emission of a photon of light. Although <i>Renilla</i> luciferase has various applications in biotechnology, its low thermal stability limits the development of its applications. Arginine is a well-known stabilizing amino acid that plays a key role in protein stabilization against inactivation. However, its impact on enzyme properties is unpredictable. This study investigates the impact of arginine on the kinetics and thermal stability of <i>Renilla</i> luciferase. The enzyme's performance was significantly enhanced in the presence of arginine, with catalytic efficiency increasing by 3.31-fold and 3.08-fold when exposed to 0.2 M and 0.3 M arginine, respectively. Additionally, arginine improved the thermal stability of <i>Renilla</i> luciferase. Molecular dynamics simulation showed that the addition of 0.2 M arginine reduced the binding of coelenteramide, the reaction product and an enzyme inhibitor, to the active site of the <i>Renilla</i> luciferase. Therefore, the release of the product was accelerated, and the affinity of <i>Renilla</i> luciferase for coelenterazine increased. Furthermore, Molecular dynamics studies indicated an increased network of water molecules surrounding <i>Renilla</i> luciferase in the presence of 0.2 M arginine. This network potentially enhances the hydrophobic effect on the protein structure, ultimately improving enzyme stability. The findings of this study hold promise for the development of commercial kits incorporating <i>Renilla</i> luciferase.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"43 4","pages":"739 - 750"},"PeriodicalIF":1.9000,"publicationDate":"2024-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Protein Journal","FirstCategoryId":"2","ListUrlMain":"https://link.springer.com/article/10.1007/s10930-024-10208-x","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Renilla luciferase catalyzes the oxidation of coelenterazine to coelenteramide and results in the emission of a photon of light. Although Renilla luciferase has various applications in biotechnology, its low thermal stability limits the development of its applications. Arginine is a well-known stabilizing amino acid that plays a key role in protein stabilization against inactivation. However, its impact on enzyme properties is unpredictable. This study investigates the impact of arginine on the kinetics and thermal stability of Renilla luciferase. The enzyme's performance was significantly enhanced in the presence of arginine, with catalytic efficiency increasing by 3.31-fold and 3.08-fold when exposed to 0.2 M and 0.3 M arginine, respectively. Additionally, arginine improved the thermal stability of Renilla luciferase. Molecular dynamics simulation showed that the addition of 0.2 M arginine reduced the binding of coelenteramide, the reaction product and an enzyme inhibitor, to the active site of the Renilla luciferase. Therefore, the release of the product was accelerated, and the affinity of Renilla luciferase for coelenterazine increased. Furthermore, Molecular dynamics studies indicated an increased network of water molecules surrounding Renilla luciferase in the presence of 0.2 M arginine. This network potentially enhances the hydrophobic effect on the protein structure, ultimately improving enzyme stability. The findings of this study hold promise for the development of commercial kits incorporating Renilla luciferase.
瑞宁拉荧光素酶催化腔肠素氧化成腔肠酰胺,从而发出光子。尽管瑞宁拉荧光素酶在生物技术中有多种应用,但其热稳定性较低,限制了其应用的发展。精氨酸是一种众所周知的稳定氨基酸,在稳定蛋白质防止失活方面起着关键作用。然而,精氨酸对酶特性的影响却难以预测。本研究探讨了精氨酸对雷尼拉荧光素酶动力学和热稳定性的影响。在精氨酸存在的情况下,酶的性能显著提高,当暴露于 0.2 M 和 0.3 M 精氨酸时,催化效率分别提高了 3.31 倍和 3.08 倍。此外,精氨酸还提高了雷尼拉荧光素酶的热稳定性。分子动力学模拟显示,加入 0.2 M 精氨酸后,反应产物和酶抑制剂腔肠酰胺与雷尼拉荧光素酶活性位点的结合减少。因此,产物的释放速度加快,而雷尼拉荧光素酶对腔肠素的亲和力增加。此外,分子动力学研究表明,在 0.2 M 精氨酸存在的情况下,Renilla 荧光素酶周围的水分子网络增加。这种网络可能会增强蛋白质结构的疏水效应,最终提高酶的稳定性。这项研究的结果为开发含有雷尼拉荧光素酶的商业试剂盒带来了希望。
期刊介绍:
The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.