A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes
Methanethiol, a gas with the characteristic smell of rotten cabbage, is a product of microbial methionine degradation. In the human body, methanethiol originates primarily from bacteria residing in the lumen of the large intestine. Selenium-binding protein 1 (SELENBP1), a marker protein of mature en...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-07-01
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| Series: | Redox Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231721001208 |
| _version_ | 1818864274985975808 |
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| author | Thilo Magnus Philipp Andreas Will Hannes Richter Patrick Rainer Winterhalter Georg Pohnert Holger Steinbrenner Lars-Oliver Klotz |
| author_facet | Thilo Magnus Philipp Andreas Will Hannes Richter Patrick Rainer Winterhalter Georg Pohnert Holger Steinbrenner Lars-Oliver Klotz |
| author_sort | Thilo Magnus Philipp |
| collection | DOAJ |
| description | Methanethiol, a gas with the characteristic smell of rotten cabbage, is a product of microbial methionine degradation. In the human body, methanethiol originates primarily from bacteria residing in the lumen of the large intestine. Selenium-binding protein 1 (SELENBP1), a marker protein of mature enterocytes, has recently been identified as a methanethiol oxidase (MTO). It catalyzes the conversion of methanethiol to hydrogen sulfide (H2S), hydrogen peroxide (H2O2) and formaldehyde. Here, human Caco-2 intestinal epithelial cells were subjected to enterocyte-like differentiation, followed by analysis of SELENBP1 levels and MTO activity. To that end, we established a novel coupled assay to assess MTO activity mimicking the proximity of microbiome and intestinal epithelial cells in vivo. The assay is based on in situ-generation of methanethiol as catalyzed by a bacterial recombinant l-methionine gamma-lyase (MGL), followed by detection of H2S and H2O2. Applying this assay, we verified the loss and impairment of MTO function in SELENBP1 variants (His329Tyr; Gly225Trp) previously identified in individuals with familial extraoral halitosis. MTO activity was strongly enhanced in Caco-2 cells upon enterocyte differentiation, in parallel with increased SELENBP1 levels. This suggests that mature enterocytes located at the tip of colonic crypts are capable of eliminating microbiome-derived methanethiol. |
| first_indexed | 2024-12-19T10:29:04Z |
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| id | doaj.art-21f8c361b9b643da9a82dd9e6bb55a8f |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-12-19T10:29:04Z |
| publishDate | 2021-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-21f8c361b9b643da9a82dd9e6bb55a8f2022-12-21T20:25:48ZengElsevierRedox Biology2213-23172021-07-0143101972A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytesThilo Magnus Philipp0Andreas Will1Hannes Richter2Patrick Rainer Winterhalter3Georg Pohnert4Holger Steinbrenner5Lars-Oliver Klotz6Institute of Nutritional Sciences, Nutrigenomics Section, Friedrich Schiller University Jena, Jena, GermanyInstitute of Nutritional Sciences, Nutrigenomics Section, Friedrich Schiller University Jena, Jena, GermanyInstitute for Inorganic and Analytical Chemistry, Friedrich Schiller University Jena, Jena, GermanyDepartment of Cardiac Surgery, Martin Luther University Halle-Wittenberg, Halle, GermanyInstitute for Inorganic and Analytical Chemistry, Friedrich Schiller University Jena, Jena, GermanyInstitute of Nutritional Sciences, Nutrigenomics Section, Friedrich Schiller University Jena, Jena, GermanyInstitute of Nutritional Sciences, Nutrigenomics Section, Friedrich Schiller University Jena, Jena, Germany; Corresponding author. Institute of Nutritional Sciences, Nutrigenomics Section, Friedrich Schiller University Jena, Dornburger Strasse 29, D-07743, Jena, Germany.Methanethiol, a gas with the characteristic smell of rotten cabbage, is a product of microbial methionine degradation. In the human body, methanethiol originates primarily from bacteria residing in the lumen of the large intestine. Selenium-binding protein 1 (SELENBP1), a marker protein of mature enterocytes, has recently been identified as a methanethiol oxidase (MTO). It catalyzes the conversion of methanethiol to hydrogen sulfide (H2S), hydrogen peroxide (H2O2) and formaldehyde. Here, human Caco-2 intestinal epithelial cells were subjected to enterocyte-like differentiation, followed by analysis of SELENBP1 levels and MTO activity. To that end, we established a novel coupled assay to assess MTO activity mimicking the proximity of microbiome and intestinal epithelial cells in vivo. The assay is based on in situ-generation of methanethiol as catalyzed by a bacterial recombinant l-methionine gamma-lyase (MGL), followed by detection of H2S and H2O2. Applying this assay, we verified the loss and impairment of MTO function in SELENBP1 variants (His329Tyr; Gly225Trp) previously identified in individuals with familial extraoral halitosis. MTO activity was strongly enhanced in Caco-2 cells upon enterocyte differentiation, in parallel with increased SELENBP1 levels. This suggests that mature enterocytes located at the tip of colonic crypts are capable of eliminating microbiome-derived methanethiol.http://www.sciencedirect.com/science/article/pii/S2213231721001208Selenium-binding protein 1Methanethiol oxidaseCaco-2 cellsHydrogen sulfideHydrogen peroxide |
| spellingShingle | Thilo Magnus Philipp Andreas Will Hannes Richter Patrick Rainer Winterhalter Georg Pohnert Holger Steinbrenner Lars-Oliver Klotz A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes Redox Biology Selenium-binding protein 1 Methanethiol oxidase Caco-2 cells Hydrogen sulfide Hydrogen peroxide |
| title | A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes |
| title_full | A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes |
| title_fullStr | A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes |
| title_full_unstemmed | A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes |
| title_short | A coupled enzyme assay for detection of selenium-binding protein 1 (SELENBP1) methanethiol oxidase (MTO) activity in mature enterocytes |
| title_sort | coupled enzyme assay for detection of selenium binding protein 1 selenbp1 methanethiol oxidase mto activity in mature enterocytes |
| topic | Selenium-binding protein 1 Methanethiol oxidase Caco-2 cells Hydrogen sulfide Hydrogen peroxide |
| url | http://www.sciencedirect.com/science/article/pii/S2213231721001208 |
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