Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury

Tetrahydrobiopterin (BH4) deficiency is caused by genetic abnormalities that impair its biosynthesis and recycling, which trigger neurochemical, metabolic, and redox imbalances. Low BH4 levels are also associated with hypoxia, reperfusion reoxygenation, endothelial dysfunction, and other conditions...

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Main Authors: R. Steven Traeger, James Woodcock, Sidhartha Tan, Zhongjie Shi, Jeannette Vasquez-Vivar
Format: Article
Language:English
Published: Elsevier 2025-02-01
Series:Redox Biology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2213231724004257
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author R. Steven Traeger
James Woodcock
Sidhartha Tan
Zhongjie Shi
Jeannette Vasquez-Vivar
author_facet R. Steven Traeger
James Woodcock
Sidhartha Tan
Zhongjie Shi
Jeannette Vasquez-Vivar
author_sort R. Steven Traeger
collection DOAJ
description Tetrahydrobiopterin (BH4) deficiency is caused by genetic abnormalities that impair its biosynthesis and recycling, which trigger neurochemical, metabolic, and redox imbalances. Low BH4 levels are also associated with hypoxia, reperfusion reoxygenation, endothelial dysfunction, and other conditions that are not genetically determined. The exact cause of changes in BH4 in nongenetic disorders is not entirely understood, but a role for oxidant species has been implicated. The oxidation of BH4 generates several products, including 7,8-dihydrobiopterin (BH2), the accumulation of which is predicted in cells with low dihydrofolate reductase activity. The relative efficiency of oxidant species at causing variations in BH4/BH2 levels in cells furnished with several antioxidant enzymes has not yet been systematically analyzed. This study examined the quantitative changes of BH4/BH2 in cells challenged with several oxidants. We showed that BH2 is not a major product of treatments with hydrogen peroxide or RSL3, as indicated by the moderate effect of dihydrofolate reductase-inhibitor methotrexate on the accumulation of BH2. However, we found a net loss in BH4/BH2, suggesting that products other than BH2 were generated. These reactions were further examined in NOX4-expressing HEK cells producing hydrogen peroxide. These cells showed slightly decreased BH4/BH2 ratios compared with HEK wild-type cells, and, again, methotrexate treatment moderately increased BH2 levels. In contrast, peroxynitrite-producing RAW 264.7 cells showed dramatically decreased BH4 levels without BH2 accumulation. Following the activation of peroxynitrite production with PMA in lipopolysaccharide-treated cells, we also found a significant time-dependent decline in GTPCH-I protein levels. We conclude that hydrogen peroxide is the least effective oxidant species at decreasing intracellular BH4 levels, while peroxynitrite is highly effective by targeting GTPCH-I and BH4 directly. Moreover, we conclude that BH4/BH2 levels are not a determinant of RSL3 cytotoxicity.
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spelling doaj-art-aaa603cebc7746f08f9f2226115f96812025-08-20T02:41:20ZengElsevierRedox Biology2213-23172025-02-017910344710.1016/j.redox.2024.103447Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injuryR. Steven Traeger0James Woodcock1Sidhartha Tan2Zhongjie Shi3Jeannette Vasquez-Vivar4Department of Biophysics and Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI, 53226, USADepartment of Biophysics and Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI, 53226, USADepartment of Pediatrics, Wayne University, Detroit, MI, 48201, USADepartment of Pediatrics, Wayne University, Detroit, MI, 48201, USADepartment of Biophysics and Redox Biology Program, Medical College of Wisconsin, Milwaukee, WI, 53226, USA; Corresponding author.Tetrahydrobiopterin (BH4) deficiency is caused by genetic abnormalities that impair its biosynthesis and recycling, which trigger neurochemical, metabolic, and redox imbalances. Low BH4 levels are also associated with hypoxia, reperfusion reoxygenation, endothelial dysfunction, and other conditions that are not genetically determined. The exact cause of changes in BH4 in nongenetic disorders is not entirely understood, but a role for oxidant species has been implicated. The oxidation of BH4 generates several products, including 7,8-dihydrobiopterin (BH2), the accumulation of which is predicted in cells with low dihydrofolate reductase activity. The relative efficiency of oxidant species at causing variations in BH4/BH2 levels in cells furnished with several antioxidant enzymes has not yet been systematically analyzed. This study examined the quantitative changes of BH4/BH2 in cells challenged with several oxidants. We showed that BH2 is not a major product of treatments with hydrogen peroxide or RSL3, as indicated by the moderate effect of dihydrofolate reductase-inhibitor methotrexate on the accumulation of BH2. However, we found a net loss in BH4/BH2, suggesting that products other than BH2 were generated. These reactions were further examined in NOX4-expressing HEK cells producing hydrogen peroxide. These cells showed slightly decreased BH4/BH2 ratios compared with HEK wild-type cells, and, again, methotrexate treatment moderately increased BH2 levels. In contrast, peroxynitrite-producing RAW 264.7 cells showed dramatically decreased BH4 levels without BH2 accumulation. Following the activation of peroxynitrite production with PMA in lipopolysaccharide-treated cells, we also found a significant time-dependent decline in GTPCH-I protein levels. We conclude that hydrogen peroxide is the least effective oxidant species at decreasing intracellular BH4 levels, while peroxynitrite is highly effective by targeting GTPCH-I and BH4 directly. Moreover, we conclude that BH4/BH2 levels are not a determinant of RSL3 cytotoxicity.http://www.sciencedirect.com/science/article/pii/S2213231724004257Hydrogen peroxidePeroxynitriteRSL3NOX4BH2Antioxidants
spellingShingle R. Steven Traeger
James Woodcock
Sidhartha Tan
Zhongjie Shi
Jeannette Vasquez-Vivar
Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
Redox Biology
Hydrogen peroxide
Peroxynitrite
RSL3
NOX4
BH2
Antioxidants
title Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
title_full Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
title_fullStr Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
title_full_unstemmed Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
title_short Tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
title_sort tetrahydrobiopterin as a rheostat of cell resistance to oxidant injury
topic Hydrogen peroxide
Peroxynitrite
RSL3
NOX4
BH2
Antioxidants
url http://www.sciencedirect.com/science/article/pii/S2213231724004257
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AT jameswoodcock tetrahydrobiopterinasarheostatofcellresistancetooxidantinjury
AT sidharthatan tetrahydrobiopterinasarheostatofcellresistancetooxidantinjury
AT zhongjieshi tetrahydrobiopterinasarheostatofcellresistancetooxidantinjury
AT jeannettevasquezvivar tetrahydrobiopterinasarheostatofcellresistancetooxidantinjury