Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1
GLUT1 is a facilitative glucose transporter that can transport oxidized vitamin C (i.e., dehydroascorbic acid) and complements the action of reduced vitamin C transporters. To identify the residues involved in human GLUT1’s transport of dehydroascorbic acid, we performed docking studies in the 5 Å g...
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MDPI AG
2023-01-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/2/521 |
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| author | Marcelo Villagrán Carlos F. Burgos Coralia I. Rivas Lorena Mardones |
| author_facet | Marcelo Villagrán Carlos F. Burgos Coralia I. Rivas Lorena Mardones |
| author_sort | Marcelo Villagrán |
| collection | DOAJ |
| description | GLUT1 is a facilitative glucose transporter that can transport oxidized vitamin C (i.e., dehydroascorbic acid) and complements the action of reduced vitamin C transporters. To identify the residues involved in human GLUT1’s transport of dehydroascorbic acid, we performed docking studies in the 5 Å grid of the glucose-binding cavity of GLUT1. The interactions of the bicyclic hemiacetal form of dehydroascorbic acid with GLUT1 through hydrogen bonds with the -OH group of C3 and C5 were less favorable than the interactions with the sugars transported by GLUT1. The eight most relevant residues in such interactions (i.e., F26, Q161, I164, Q282, Y292, and W412) were mutated to alanine to perform functional studies for dehydroascorbic acid and the glucose analog, 2-deoxiglucose, in <i>Xenopus laevis</i> oocytes. All the mutants decreased the uptake of both substrates to less than 50%. The partial effect of the N317A mutant in transporting dehydroascorbic acid was associated with a 30% decrease in the V<sub>max</sub> compared to the wildtype GLUT1. The results show that both substrates share the eight residues studied in GLUT1, albeit with a differential contribution of N317. Our work, combining docking with functional studies, marks the first to identify structural determinants of oxidized vitamin C’s transport via GLUT1. |
| first_indexed | 2024-03-09T11:37:18Z |
| format | Article |
| id | doaj.art-9448f7d4c0e8485cb62297b2616ef696 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T11:37:18Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-9448f7d4c0e8485cb62297b2616ef6962023-11-30T23:41:01ZengMDPI AGMolecules1420-30492023-01-0128252110.3390/molecules28020521Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1Marcelo Villagrán0Carlos F. Burgos1Coralia I. Rivas2Lorena Mardones3Research Laboratory in Biological Sciences, Department of Basic Sciences, Medicine Faculty, Universidad Católica de la Santísima Concepción, Concepción 4090541, ChileLaboratory of Neurophysiology, Department of Physiology, Faculty of Biological Sciences, Universidad de Concepción, Concepción 4030000, ChileLaboratory of Antioxidants, Department of Pathophysiology, Faculty of Biological Sciences, Universidad de Concepción, Concepción 4070386, ChileResearch Laboratory in Biological Sciences, Department of Basic Sciences, Medicine Faculty, Universidad Católica de la Santísima Concepción, Concepción 4090541, ChileGLUT1 is a facilitative glucose transporter that can transport oxidized vitamin C (i.e., dehydroascorbic acid) and complements the action of reduced vitamin C transporters. To identify the residues involved in human GLUT1’s transport of dehydroascorbic acid, we performed docking studies in the 5 Å grid of the glucose-binding cavity of GLUT1. The interactions of the bicyclic hemiacetal form of dehydroascorbic acid with GLUT1 through hydrogen bonds with the -OH group of C3 and C5 were less favorable than the interactions with the sugars transported by GLUT1. The eight most relevant residues in such interactions (i.e., F26, Q161, I164, Q282, Y292, and W412) were mutated to alanine to perform functional studies for dehydroascorbic acid and the glucose analog, 2-deoxiglucose, in <i>Xenopus laevis</i> oocytes. All the mutants decreased the uptake of both substrates to less than 50%. The partial effect of the N317A mutant in transporting dehydroascorbic acid was associated with a 30% decrease in the V<sub>max</sub> compared to the wildtype GLUT1. The results show that both substrates share the eight residues studied in GLUT1, albeit with a differential contribution of N317. Our work, combining docking with functional studies, marks the first to identify structural determinants of oxidized vitamin C’s transport via GLUT1.https://www.mdpi.com/1420-3049/28/2/521GLUT1glucose transporterdehydroascorbic acid |
| spellingShingle | Marcelo Villagrán Carlos F. Burgos Coralia I. Rivas Lorena Mardones Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 Molecules GLUT1 glucose transporter dehydroascorbic acid |
| title | Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 |
| title_full | Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 |
| title_fullStr | Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 |
| title_full_unstemmed | Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 |
| title_short | Identification of Structural Determinants of the Transport of the Dehydroascorbic Acid Mediated by Glucose Transport GLUT1 |
| title_sort | identification of structural determinants of the transport of the dehydroascorbic acid mediated by glucose transport glut1 |
| topic | GLUT1 glucose transporter dehydroascorbic acid |
| url | https://www.mdpi.com/1420-3049/28/2/521 |
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