Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase
Vitamin K epoxide reductase (VKOR) is a target enzyme for anticoagulants, such as warfarin, that are used as medicines or rodenticides. Assessing VKOR activity is required to ensure the proper usage of these drugs. Dithiothreitol (DTT) is a typical disulfide reductant that is used as a substrate for...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Veterinary and Animal Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2451943X20300089 |
| _version_ | 1818257031439581184 |
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| author | Kazuki Takeda Ayuko Morita Yoshinori Ikenaka Shouta M.M. Nakayama Mayumi Ishizuka |
| author_facet | Kazuki Takeda Ayuko Morita Yoshinori Ikenaka Shouta M.M. Nakayama Mayumi Ishizuka |
| author_sort | Kazuki Takeda |
| collection | DOAJ |
| description | Vitamin K epoxide reductase (VKOR) is a target enzyme for anticoagulants, such as warfarin, that are used as medicines or rodenticides. Assessing VKOR activity is required to ensure the proper usage of these drugs. Dithiothreitol (DTT) is a typical disulfide reductant that is used as a substrate for in vitro VKOR assays. However, DTT is considered problematic because of its side effects. Tris(3-hydroxypropyl)phosphine (THP) has been found to be a reliable alternative to DTT, as shown by kinetic analyses of the VKOR with them. THP showed significantly lower Vmax and Km values than those of DTT; however, there was no significant difference in their Vmax/Km and IC50 for warfarin. |
| first_indexed | 2024-12-12T17:37:11Z |
| format | Article |
| id | doaj.art-3421d9b867c94d65bd9d2eb8c5b8b934 |
| institution | Directory Open Access Journal |
| issn | 2451-943X |
| language | English |
| last_indexed | 2024-12-12T17:37:11Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Veterinary and Animal Science |
| spelling | doaj.art-3421d9b867c94d65bd9d2eb8c5b8b9342022-12-22T00:17:11ZengElsevierVeterinary and Animal Science2451-943X2020-06-019100095Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductaseKazuki Takeda0Ayuko Morita1Yoshinori Ikenaka2Shouta M.M. Nakayama3Mayumi Ishizuka4Department of Environmental Sciences, Laboratory of Toxicology, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo 060-0818, JapanDepartment of Environmental Sciences, Laboratory of Toxicology, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo 060-0818, JapanDepartment of Environmental Sciences, Laboratory of Toxicology, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo 060-0818, Japan; Water Research Group, Unit for Environmental Sciences and Management, North-West University, Potchefstroom, South AfricaDepartment of Environmental Sciences, Laboratory of Toxicology, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo 060-0818, JapanDepartment of Environmental Sciences, Laboratory of Toxicology, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo 060-0818, Japan; Corresponding author.Vitamin K epoxide reductase (VKOR) is a target enzyme for anticoagulants, such as warfarin, that are used as medicines or rodenticides. Assessing VKOR activity is required to ensure the proper usage of these drugs. Dithiothreitol (DTT) is a typical disulfide reductant that is used as a substrate for in vitro VKOR assays. However, DTT is considered problematic because of its side effects. Tris(3-hydroxypropyl)phosphine (THP) has been found to be a reliable alternative to DTT, as shown by kinetic analyses of the VKOR with them. THP showed significantly lower Vmax and Km values than those of DTT; however, there was no significant difference in their Vmax/Km and IC50 for warfarin.http://www.sciencedirect.com/science/article/pii/S2451943X20300089Vitamin KVitamin K epoxide reductaseDithiothreitolTris(3-hydroxypropyl)phosphineanticoagulant |
| spellingShingle | Kazuki Takeda Ayuko Morita Yoshinori Ikenaka Shouta M.M. Nakayama Mayumi Ishizuka Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase Veterinary and Animal Science Vitamin K Vitamin K epoxide reductase Dithiothreitol Tris(3-hydroxypropyl)phosphine anticoagulant |
| title | Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase |
| title_full | Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase |
| title_fullStr | Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase |
| title_full_unstemmed | Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase |
| title_short | Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase |
| title_sort | comparison of two reducing agents dithiothreitol and tris 3 hydroxypropyl phosphine for in vitro kinetic assay of vitamin k epoxide reductase |
| topic | Vitamin K Vitamin K epoxide reductase Dithiothreitol Tris(3-hydroxypropyl)phosphine anticoagulant |
| url | http://www.sciencedirect.com/science/article/pii/S2451943X20300089 |
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