The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice
Legumain is a lysosomal cysteine protease that has been implicated in an increasing amount of physiological and pathophysiological processes. However, the upstream mechanisms regulating the expression and function of legumain are not well understood. Here, we provide in vitro and in vivo data showin...
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MDPI AG
2023-12-01
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| Online Access: | https://www.mdpi.com/2073-4409/13/1/36 |
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| author | Karl Martin Forbord Meshail Okla Ngoc Nguyen Lunde Tatjana Bosnjak-Olsen Guro Arnekleiv Daniel Hesselson Harald Thidemann Johansen Jonathan C. Y. Tang Moustapha Kassem Rigmor Solberg Abbas Jafari |
| author_facet | Karl Martin Forbord Meshail Okla Ngoc Nguyen Lunde Tatjana Bosnjak-Olsen Guro Arnekleiv Daniel Hesselson Harald Thidemann Johansen Jonathan C. Y. Tang Moustapha Kassem Rigmor Solberg Abbas Jafari |
| author_sort | Karl Martin Forbord |
| collection | DOAJ |
| description | Legumain is a lysosomal cysteine protease that has been implicated in an increasing amount of physiological and pathophysiological processes. However, the upstream mechanisms regulating the expression and function of legumain are not well understood. Here, we provide in vitro and in vivo data showing that vitamin D<sub>3</sub> (VD<sub>3</sub>) enhances legumain expression and function. In turn, legumain alters VD<sub>3</sub> bioavailability, possibly through proteolytic cleavage of vitamin D binding protein (VDBP). Active VD<sub>3</sub> (1,25(OH)<sub>2</sub>D<sub>3</sub>) increased legumain expression, activity, and secretion in osteogenic cultures of human bone marrow stromal cells. Upregulation of legumain was also observed in vivo, evidenced by increased legumain mRNA in the liver and spleen, as well as increased legumain activity in kidneys from wild-type mice treated with 25(OH)D<sub>3</sub> (50 µg/kg, subcutaneously) for 8 days compared to a control. In addition, the serum level of legumain was also increased. We further showed that active legumain cleaved purified VDBP (55 kDa) in vitro, forming a 45 kDa fragment. In vivo, no VDBP cleavage was found in kidneys or liver from legumain-deficient mice (<i>Lgmn</i><sup>−/−</sup>), whereas VDBP was cleaved in wild-type control mice (<i>Lgmn</i><sup>+/+</sup>). Finally, legumain deficiency resulted in increased plasma levels of 25(OH)D<sub>3</sub> and total VD<sub>3</sub> and altered expression of key renal enzymes involved in VD<sub>3</sub> metabolism (CYP24A1 and CYP27B1). In conclusion, a regulatory interplay between VD<sub>3</sub> and legumain is suggested. |
| first_indexed | 2024-03-08T15:10:42Z |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-08T15:10:42Z |
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| spelling | doaj.art-bdf2c9a035db4a4e847885aa1093ff8a2024-01-10T14:53:16ZengMDPI AGCells2073-44092023-12-011313610.3390/cells13010036The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in MiceKarl Martin Forbord0Meshail Okla1Ngoc Nguyen Lunde2Tatjana Bosnjak-Olsen3Guro Arnekleiv4Daniel Hesselson5Harald Thidemann Johansen6Jonathan C. Y. Tang7Moustapha Kassem8Rigmor Solberg9Abbas Jafari10Section for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwayDepartment of Endocrinology and Metabolism, Odense University Hospital, University of Southern Denmark, 5230 Odense, DenmarkSection for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwaySection for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwaySection for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwayCentenary Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2006, AustraliaSection for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwayBioanalytical Facility, Norwich Medical School, University of East Anglia, Norwich NR4 7TJ, UKDepartment of Endocrinology and Metabolism, Odense University Hospital, University of Southern Denmark, 5230 Odense, DenmarkSection for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, NorwayDepartment of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, DenmarkLegumain is a lysosomal cysteine protease that has been implicated in an increasing amount of physiological and pathophysiological processes. However, the upstream mechanisms regulating the expression and function of legumain are not well understood. Here, we provide in vitro and in vivo data showing that vitamin D<sub>3</sub> (VD<sub>3</sub>) enhances legumain expression and function. In turn, legumain alters VD<sub>3</sub> bioavailability, possibly through proteolytic cleavage of vitamin D binding protein (VDBP). Active VD<sub>3</sub> (1,25(OH)<sub>2</sub>D<sub>3</sub>) increased legumain expression, activity, and secretion in osteogenic cultures of human bone marrow stromal cells. Upregulation of legumain was also observed in vivo, evidenced by increased legumain mRNA in the liver and spleen, as well as increased legumain activity in kidneys from wild-type mice treated with 25(OH)D<sub>3</sub> (50 µg/kg, subcutaneously) for 8 days compared to a control. In addition, the serum level of legumain was also increased. We further showed that active legumain cleaved purified VDBP (55 kDa) in vitro, forming a 45 kDa fragment. In vivo, no VDBP cleavage was found in kidneys or liver from legumain-deficient mice (<i>Lgmn</i><sup>−/−</sup>), whereas VDBP was cleaved in wild-type control mice (<i>Lgmn</i><sup>+/+</sup>). Finally, legumain deficiency resulted in increased plasma levels of 25(OH)D<sub>3</sub> and total VD<sub>3</sub> and altered expression of key renal enzymes involved in VD<sub>3</sub> metabolism (CYP24A1 and CYP27B1). In conclusion, a regulatory interplay between VD<sub>3</sub> and legumain is suggested.https://www.mdpi.com/2073-4409/13/1/36asparaginyl endopeptidaselegumainmetabolismproteolysisvitamin D |
| spellingShingle | Karl Martin Forbord Meshail Okla Ngoc Nguyen Lunde Tatjana Bosnjak-Olsen Guro Arnekleiv Daniel Hesselson Harald Thidemann Johansen Jonathan C. Y. Tang Moustapha Kassem Rigmor Solberg Abbas Jafari The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice Cells asparaginyl endopeptidase legumain metabolism proteolysis vitamin D |
| title | The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice |
| title_full | The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice |
| title_fullStr | The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice |
| title_full_unstemmed | The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice |
| title_short | The Cysteine Protease Legumain Is Upregulated by Vitamin D and Is a Regulator of Vitamin D Metabolism in Mice |
| title_sort | cysteine protease legumain is upregulated by vitamin d and is a regulator of vitamin d metabolism in mice |
| topic | asparaginyl endopeptidase legumain metabolism proteolysis vitamin D |
| url | https://www.mdpi.com/2073-4409/13/1/36 |
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