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It is unknown whether ascorbate alone (vitamin C), its oxidized
metabolite dehydroascorbic acid alone, or both species are transported
into human cells. This problem was addressed using specific assays for
each compound, freshly synthesized pure dehydroascorbic acid, the
specially synthesized analog 6-chloroascorbate, and a new assay for
6-chloroascorbate. Ascorbate and dehydroascorbic acid were transported
and accumulated distinctly; neither competed with the other. Ascorbate
was accumulated as ascorbate by sodium-dependent carrier-mediated
active transport. Dehydroascorbic acid transport and accumulation as
ascorbate was at least 10-fold faster than ascorbate transport and was
sodium-independent. Once transported, dehydroascorbic acid was
immediately reduced intracellularly to ascorbate. The analog
6-chloroascorbate had no effect on dehydroascorbic acid transport but
was a competitive inhibitor of ascorbate transport. The
K
Volume 270,
Number 21,
Issue of May 26, pp. 12584-12592, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
for 6-chloroascorbate (2.9-4.4
µM) was similar to the K
for
ascorbate transport (9.8-12.6 µM). 6-Chloroascorbate
was itself transported and accumulated in fibroblasts by a
sodium-dependent transporter. These data provide new information that
ascorbate and dehydroascorbic acid are transported into human
neutrophils and fibroblasts by two distinct mechanisms and that the
compound available for intracellular utilization is ascorbate.
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