Redox modulation of thimet oligopeptidase activity by hydrogen peroxide
Thimet oligopeptidase (EC 3.4.24.15, TOP) is a cytosolic mammalian zinc protease that can process a diversity of bioactive peptides. TOP has been pointed out as one of the main postproteasomal enzymes that process peptide antigens in the MHC class I presentation route. In the present study, we descr...
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| Format: | Article |
| Language: | English |
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Wiley
2017-07-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.12245 |
| _version_ | 1798018861877952512 |
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| author | Marcelo Y. Icimoto Juliana C. Ferreira César H. Yokomizo Larissa V. Bim Alyne Marem Joyce M. Gilio Vitor Oliveira Iseli L. Nantes |
| author_facet | Marcelo Y. Icimoto Juliana C. Ferreira César H. Yokomizo Larissa V. Bim Alyne Marem Joyce M. Gilio Vitor Oliveira Iseli L. Nantes |
| author_sort | Marcelo Y. Icimoto |
| collection | DOAJ |
| description | Thimet oligopeptidase (EC 3.4.24.15, TOP) is a cytosolic mammalian zinc protease that can process a diversity of bioactive peptides. TOP has been pointed out as one of the main postproteasomal enzymes that process peptide antigens in the MHC class I presentation route. In the present study, we describe a fine regulation of TOP activity by hydrogen peroxide (H2O2). Cells from a human embryonic kidney cell line (HEK293) underwent an ischemia/reoxygenation‐like condition known to increase H2O2 production. Immediately after reoxygenation, HEK293 cells exhibited a 32% increase in TOP activity, but no TOP activity was observed 2 h after reoxygenation. In another model, recombinant rat TOP (rTOP) was challenged by H2O2 produced by rat liver mitoplasts (RLMt) alone, and in combination with antimycin A, succinate, and antimycin A plus succinate. In these conditions, rTOP activity increased 17, 30, 32 and 38%, respectively. Determination of H2O2 concentration generated in reoxygenated cells and mitoplasts suggested a possible modulation of rTOP activity dependent on the concentration of H2O2. The measure of pure rTOP activity as a function of H2O2 concentration corroborated this hypothesis. The data fitted to an asymmetrical bell‐shaped curve in which the optimal activating H2O2 concentration was 1.2 nM, and the maximal inhibition (75% about the control) was 1 μm. Contrary to the oxidation produced by aging associated with enzyme oligomerization and inhibition, H2O2 oxidation produced sulfenic acid and maintained rTOP in the monomeric form. Consistent with the involvement of rTOP in a signaling redox cascade, the H2O2‐oxidized rTOP reacted with dimeric thioredoxin‐1 (TRx‐1) and remained covalently bound to one subunit of TRx‐1. |
| first_indexed | 2024-04-11T16:31:12Z |
| format | Article |
| id | doaj.art-34abc682f8d248868a1ff9094f7ddafd |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-11T16:31:12Z |
| publishDate | 2017-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-34abc682f8d248868a1ff9094f7ddafd2022-12-22T04:14:01ZengWileyFEBS Open Bio2211-54632017-07-01771037105010.1002/2211-5463.12245Redox modulation of thimet oligopeptidase activity by hydrogen peroxideMarcelo Y. Icimoto0Juliana C. Ferreira1César H. Yokomizo2Larissa V. Bim3Alyne Marem4Joyce M. Gilio5Vitor Oliveira6Iseli L. Nantes7Departamento de Biofísica Universidade Federal de São Paulo BrazilLaboratório de Nanoestruturas para Biologia e Materiais Avançados Centro de Ciências Naturais e Humanas Universidade Federal do ABC Santo André BrazilLaboratório de Nanoestruturas para Biologia e Materiais Avançados Centro de Ciências Naturais e Humanas Universidade Federal do ABC Santo André BrazilDepartamento de Biofísica Universidade Federal de São Paulo BrazilDepartamento de Biofísica Universidade Federal de São Paulo BrazilDepartamento de Biofísica Universidade Federal de São Paulo BrazilDepartamento de Biofísica Universidade Federal de São Paulo BrazilLaboratório de Nanoestruturas para Biologia e Materiais Avançados Centro de Ciências Naturais e Humanas Universidade Federal do ABC Santo André BrazilThimet oligopeptidase (EC 3.4.24.15, TOP) is a cytosolic mammalian zinc protease that can process a diversity of bioactive peptides. TOP has been pointed out as one of the main postproteasomal enzymes that process peptide antigens in the MHC class I presentation route. In the present study, we describe a fine regulation of TOP activity by hydrogen peroxide (H2O2). Cells from a human embryonic kidney cell line (HEK293) underwent an ischemia/reoxygenation‐like condition known to increase H2O2 production. Immediately after reoxygenation, HEK293 cells exhibited a 32% increase in TOP activity, but no TOP activity was observed 2 h after reoxygenation. In another model, recombinant rat TOP (rTOP) was challenged by H2O2 produced by rat liver mitoplasts (RLMt) alone, and in combination with antimycin A, succinate, and antimycin A plus succinate. In these conditions, rTOP activity increased 17, 30, 32 and 38%, respectively. Determination of H2O2 concentration generated in reoxygenated cells and mitoplasts suggested a possible modulation of rTOP activity dependent on the concentration of H2O2. The measure of pure rTOP activity as a function of H2O2 concentration corroborated this hypothesis. The data fitted to an asymmetrical bell‐shaped curve in which the optimal activating H2O2 concentration was 1.2 nM, and the maximal inhibition (75% about the control) was 1 μm. Contrary to the oxidation produced by aging associated with enzyme oligomerization and inhibition, H2O2 oxidation produced sulfenic acid and maintained rTOP in the monomeric form. Consistent with the involvement of rTOP in a signaling redox cascade, the H2O2‐oxidized rTOP reacted with dimeric thioredoxin‐1 (TRx‐1) and remained covalently bound to one subunit of TRx‐1.https://doi.org/10.1002/2211-5463.12245EC 3.4.24.15hydrogen peroxideprotein oxidationthioredoxin |
| spellingShingle | Marcelo Y. Icimoto Juliana C. Ferreira César H. Yokomizo Larissa V. Bim Alyne Marem Joyce M. Gilio Vitor Oliveira Iseli L. Nantes Redox modulation of thimet oligopeptidase activity by hydrogen peroxide FEBS Open Bio EC 3.4.24.15 hydrogen peroxide protein oxidation thioredoxin |
| title | Redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| title_full | Redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| title_fullStr | Redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| title_full_unstemmed | Redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| title_short | Redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| title_sort | redox modulation of thimet oligopeptidase activity by hydrogen peroxide |
| topic | EC 3.4.24.15 hydrogen peroxide protein oxidation thioredoxin |
| url | https://doi.org/10.1002/2211-5463.12245 |
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