Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection
Various chemical probes for the detection of reactive oxygen species have been developed to examine oxidative stress associated with different pathologies. L-012, a luminol-based chemiluminescent probe, is widely used to detect extracellular superoxide because of its high sensitivity. We herein demo...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-12-01
|
| Series: | Frontiers in Medicine |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmed.2022.941180/full |
| _version_ | 1798003902184947712 |
|---|---|
| author | Misaki Matsumoto Hikari Sawada Kazumi Iwata Masakazu Ibi Nozomi Asaoka Masato Katsuyama Kaori Shintani-Ishida Hiroshi Ikegaya Shigehiko Takegami Atsushi Umemura Chihiro Yabe-Nishimura |
| author_facet | Misaki Matsumoto Hikari Sawada Kazumi Iwata Masakazu Ibi Nozomi Asaoka Masato Katsuyama Kaori Shintani-Ishida Hiroshi Ikegaya Shigehiko Takegami Atsushi Umemura Chihiro Yabe-Nishimura |
| author_sort | Misaki Matsumoto |
| collection | DOAJ |
| description | Various chemical probes for the detection of reactive oxygen species have been developed to examine oxidative stress associated with different pathologies. L-012, a luminol-based chemiluminescent probe, is widely used to detect extracellular superoxide because of its high sensitivity. We herein demonstrated that the co-application of the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence without affecting the background. More than a 5-fold increase was detected in the total luminescence of L-012 in both NADPH oxidase-expressing cells and the xanthine oxidase-dependent cell-free superoxide generation system, but not in their background. Therefore, bortezomib increased the signal-to-background ratio and improved the detection of low levels of superoxide. The application of MLN2238, another peptide boronic acid proteasome inhibitor, also enhanced the luminescence of L-012. In contrast, carfilzomib, an epoxyketone proteasome inhibitor, did not increase luminescence, suggesting that the effects of bortezomib depend on the chemical structure of the peptide boronic acid, but not on its pharmacological effects. Bortezomib-induced enhancements appeared to be specific to the detection of superoxide because the detection of H2O2 by Amplex Red/HRP was not affected by the application of bortezomib. In the quantitative detection of the superoxide-specific oxidative product 2-hydroxyethidium (2-OH-E+), the application of bortezomib resulted in a 2-fold increase in the level of 2-OH-E+. Therefore, bortezomib sensitizes the detection of superoxide in both cell-based and cell-free systems, highlighting a novel feature of compounds containing the peptide boronic acid as powerful enhancers for the detection of superoxide. |
| first_indexed | 2024-04-11T12:15:08Z |
| format | Article |
| id | doaj.art-99bf2808f62a401293b176f241bd5acf |
| institution | Directory Open Access Journal |
| issn | 2296-858X |
| language | English |
| last_indexed | 2024-04-11T12:15:08Z |
| publishDate | 2022-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Medicine |
| spelling | doaj.art-99bf2808f62a401293b176f241bd5acf2022-12-22T04:24:21ZengFrontiers Media S.A.Frontiers in Medicine2296-858X2022-12-01910.3389/fmed.2022.941180941180Bortezomib is an effective enhancer for chemical probe-dependent superoxide detectionMisaki Matsumoto0Hikari Sawada1Kazumi Iwata2Masakazu Ibi3Nozomi Asaoka4Masato Katsuyama5Kaori Shintani-Ishida6Hiroshi Ikegaya7Shigehiko Takegami8Atsushi Umemura9Chihiro Yabe-Nishimura10Department of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanRadioisotope Center, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Forensic Medicine, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Forensic Medicine, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Analytical Chemistry, Kyoto Pharmaceutical University, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanDepartment of Pharmacology, Kyoto Prefectural University of Medicine, Kyoto, JapanVarious chemical probes for the detection of reactive oxygen species have been developed to examine oxidative stress associated with different pathologies. L-012, a luminol-based chemiluminescent probe, is widely used to detect extracellular superoxide because of its high sensitivity. We herein demonstrated that the co-application of the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence without affecting the background. More than a 5-fold increase was detected in the total luminescence of L-012 in both NADPH oxidase-expressing cells and the xanthine oxidase-dependent cell-free superoxide generation system, but not in their background. Therefore, bortezomib increased the signal-to-background ratio and improved the detection of low levels of superoxide. The application of MLN2238, another peptide boronic acid proteasome inhibitor, also enhanced the luminescence of L-012. In contrast, carfilzomib, an epoxyketone proteasome inhibitor, did not increase luminescence, suggesting that the effects of bortezomib depend on the chemical structure of the peptide boronic acid, but not on its pharmacological effects. Bortezomib-induced enhancements appeared to be specific to the detection of superoxide because the detection of H2O2 by Amplex Red/HRP was not affected by the application of bortezomib. In the quantitative detection of the superoxide-specific oxidative product 2-hydroxyethidium (2-OH-E+), the application of bortezomib resulted in a 2-fold increase in the level of 2-OH-E+. Therefore, bortezomib sensitizes the detection of superoxide in both cell-based and cell-free systems, highlighting a novel feature of compounds containing the peptide boronic acid as powerful enhancers for the detection of superoxide.https://www.frontiersin.org/articles/10.3389/fmed.2022.941180/fullbortezomibsuperoxideNADPH oxidasexanthine oxidasepeptide boronic acid |
| spellingShingle | Misaki Matsumoto Hikari Sawada Kazumi Iwata Masakazu Ibi Nozomi Asaoka Masato Katsuyama Kaori Shintani-Ishida Hiroshi Ikegaya Shigehiko Takegami Atsushi Umemura Chihiro Yabe-Nishimura Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection Frontiers in Medicine bortezomib superoxide NADPH oxidase xanthine oxidase peptide boronic acid |
| title | Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection |
| title_full | Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection |
| title_fullStr | Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection |
| title_full_unstemmed | Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection |
| title_short | Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection |
| title_sort | bortezomib is an effective enhancer for chemical probe dependent superoxide detection |
| topic | bortezomib superoxide NADPH oxidase xanthine oxidase peptide boronic acid |
| url | https://www.frontiersin.org/articles/10.3389/fmed.2022.941180/full |
| work_keys_str_mv | AT misakimatsumoto bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT hikarisawada bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT kazumiiwata bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT masakazuibi bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT nozomiasaoka bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT masatokatsuyama bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT kaorishintaniishida bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT hiroshiikegaya bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT shigehikotakegami bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT atsushiumemura bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection AT chihiroyabenishimura bortezomibisaneffectiveenhancerforchemicalprobedependentsuperoxidedetection |