Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies
Renilla luciferase catalyzes the oxidation of coelenterazine to coelenteramide, resulting in the emission of a photon of light. This study investigated the impact of sorbitol on the structural and kinetic properties of Renilla luciferase using circular dichroism, fluorescence spectroscopy, and molec...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | Biochemistry and Biophysics Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S240558082300198X |
| _version_ | 1827353650109874176 |
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| author | Golnoosh Khoshnevisan Rahman Emamzadeh Mahboobeh Nazari Mina Oliayi Reyhaneh Sariri |
| author_facet | Golnoosh Khoshnevisan Rahman Emamzadeh Mahboobeh Nazari Mina Oliayi Reyhaneh Sariri |
| author_sort | Golnoosh Khoshnevisan |
| collection | DOAJ |
| description | Renilla luciferase catalyzes the oxidation of coelenterazine to coelenteramide, resulting in the emission of a photon of light. This study investigated the impact of sorbitol on the structural and kinetic properties of Renilla luciferase using circular dichroism, fluorescence spectroscopy, and molecular dynamics simulations. Our investigation, carried out using circular dichroism and fluorescence analyses, as well as a thermal stability assay, has revealed that sorbitol induces conformational changes in the enzyme but does not improve its thermal stability. Moreover, through kinetic studies, it has been demonstrated that at a concentration of 0.4 M, sorbitol enhances the catalytic efficiency of Renilla luciferase. However, at higher concentrations, sorbitol results in a decrease in catalytic efficiency. Additionally, molecular dynamics simulations have shown that sorbitol increases the presence of hydrophobic pockets on the enzyme's surface. These simulations have also provided evidence that at a concentration of 0.4 M, sorbitol facilitates substrate access to the active site of the enzyme. Nevertheless, at higher concentrations, sorbitol obstructs substrate trafficking, most likely due to its impact on the gateway to the active site. This study may provide insights into the kinetic changes observed in enzymes with buried active sites, such as those with α/β hydrolase fold. |
| first_indexed | 2024-03-08T03:30:31Z |
| format | Article |
| id | doaj.art-a3b717bb1d5c49c380024eaee941afd6 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-03-08T03:30:31Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-a3b717bb1d5c49c380024eaee941afd62024-02-11T05:11:16ZengElsevierBiochemistry and Biophysics Reports2405-58082024-03-0137101617Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studiesGolnoosh Khoshnevisan0Rahman Emamzadeh1Mahboobeh Nazari2Mina Oliayi3Reyhaneh Sariri4Department of Biology, University of Guilan, Rasht, IranDepartment of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran; Corresponding author.Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences, Tehran, Iran; Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, IranDepartment of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, IranDepartment of Biology, University of Guilan, Rasht, IranRenilla luciferase catalyzes the oxidation of coelenterazine to coelenteramide, resulting in the emission of a photon of light. This study investigated the impact of sorbitol on the structural and kinetic properties of Renilla luciferase using circular dichroism, fluorescence spectroscopy, and molecular dynamics simulations. Our investigation, carried out using circular dichroism and fluorescence analyses, as well as a thermal stability assay, has revealed that sorbitol induces conformational changes in the enzyme but does not improve its thermal stability. Moreover, through kinetic studies, it has been demonstrated that at a concentration of 0.4 M, sorbitol enhances the catalytic efficiency of Renilla luciferase. However, at higher concentrations, sorbitol results in a decrease in catalytic efficiency. Additionally, molecular dynamics simulations have shown that sorbitol increases the presence of hydrophobic pockets on the enzyme's surface. These simulations have also provided evidence that at a concentration of 0.4 M, sorbitol facilitates substrate access to the active site of the enzyme. Nevertheless, at higher concentrations, sorbitol obstructs substrate trafficking, most likely due to its impact on the gateway to the active site. This study may provide insights into the kinetic changes observed in enzymes with buried active sites, such as those with α/β hydrolase fold.http://www.sciencedirect.com/science/article/pii/S240558082300198XRenilla luciferaseSorbitolEnzyme kineticsMolecular dynamicsCatalytic efficiency |
| spellingShingle | Golnoosh Khoshnevisan Rahman Emamzadeh Mahboobeh Nazari Mina Oliayi Reyhaneh Sariri Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies Biochemistry and Biophysics Reports Renilla luciferase Sorbitol Enzyme kinetics Molecular dynamics Catalytic efficiency |
| title | Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies |
| title_full | Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies |
| title_fullStr | Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies |
| title_full_unstemmed | Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies |
| title_short | Uncovering the role of sorbitol in Renilla luciferase kinetics: Insights from spectroscopic and molecular dynamics studies |
| title_sort | uncovering the role of sorbitol in renilla luciferase kinetics insights from spectroscopic and molecular dynamics studies |
| topic | Renilla luciferase Sorbitol Enzyme kinetics Molecular dynamics Catalytic efficiency |
| url | http://www.sciencedirect.com/science/article/pii/S240558082300198X |
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