The dimer of human SVCT1 is key for transport function
File(s)
Published version
Author(s)
Woubshete, Menebere
Chan, Lok I
Diallinas, George
Byrne, Bernadette
Type
Journal Article
Abstract
Humans and other primates lack the ability to synthesize the essential nutrient, Vitamin C, which is derived exclusively from the diet. Crucial for effective vitamin C uptake are the Na+ dependent Vitamin C transporters, SVCT1 and SVCT2, members of the nucleobase ascorbate transporter (NAT) family. SVCT1 and 2 actively transport the reduced form of Vitamin C, ascorbic acid, into key tissues. The recent structure of the mouse SVCT1 revealed the molecular basis of substrate binding and that, like the other structurally characterised members of the NAT family, it exists as a closely associated dimer. SVCT1 is likely to function via the elevator mechanism with the core domain of each protomer able to bind substrate and move through the membrane carrying the substrate across the membrane. Here we explored the function of a range of variants of the human SVCT1, revealing a range of residues involved in substrate selection and binding, and confirming the importance of the C-terminus in membrane localisation. Furthermore, using a dominant negative mutant we show that the dimer is essential for transport function, as previously seen in the fungal homologue, UapA. In addition, we show that a localisation deficient C-terminal truncation of SVCT1 blocks correct localisation of co-expressed, associated wildtype SVCT1. These results clearly show the importance of the dimer in both correct SVCT1 trafficking and transport activity.
Date Issued
2024-12-01
Date Acceptance
2024-10-03
Citation
Biochimica et Biophysica Acta - Biomembranes, 2024, 1866 (8)
ISSN
0005-2736
Publisher
Elsevier
Journal / Book Title
Biochimica et Biophysica Acta - Biomembranes
Volume
1866
Issue
8
Copyright Statement
© 2024 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/39369805
PII: S0005-2736(24)00121-4
Subjects
ASCORBATE
Biochemistry & Molecular Biology
Biophysics
Dimerisation
ER-EXIT
FAMILY
Life Sciences & Biomedicine
LOCALIZATION
MECHANISTIC INSIGHTS
Mutants
N-GLYCOSYLATION
PURINE TRANSPORTER
Science & Technology
Substrate uptake
SVCT1
Transport
UAPA
URIC-ACID
Vitamin C
VITAMIN-C TRANSPORTER
Publication Status
Published
Coverage Spatial
Netherlands
Article Number
184390
Date Publish Online
2024-10-04