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Ascorbate uptake in pig coronary artery endothelial cells

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Abstract

Although smooth muscle and endothelial cells in pig coronary artery are morphologically and functionally distinct, ascorbate uptake has been characterized only in smooth muscle cells. Ascorbate transporters in kidney and intestinal epithelial cells differ from those in smooth muscle. We examined ascorbate transport and mRNA expression of sodium-dependent vitamin C transporters (SVCT) by RT-PCR in the pig coronary artery endothelial cell cultures. When 14C-ascorbate uptake in endothelial cells was examined as 14C or by HPLC, the two values did not differ from each other. 14C-ascorbate uptake was Na+-dependent, stereoselective for l-ascorbate and inhibited by sulfinpyrazone. The kinetic characteristics of the uptake were: Km = 27± 3 μM (Hill coefficient = 1) for ascorbate and Km = 73± 14 mM (Hill coefficient = 2) for Na+. Surprisingly, endothelial cells had similar kinetic parameters as smooth muscle cells, except for a slightly lower uptake velocity in endothelial cells. Comparison with the smooth muscle showed that both tissue types expressed mRNA for SVCT2. Endothelial cells differ from epithelial cells which express mainly SVCT1 but resemble smooth muscle cells in this respect. (Mol Cell Biochem 271: 43–49, 2005)

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Abbreviations

DHAA:

dehydroascorbic acid

DTT:

dithiothreitol

G3PDH:

glyceraldehyde 3-phosphate dehydrogenase

HEPES:

4-(2-hydroxyethyl-1-poperazine ethane sulfonate

NMG:

N-methyl-glucamine+

PCEC:

pig coronary artery endothelial cells

PCSMC:

pig coronary artery smooth muscle cells

SERCA:

sarco/endoplasmic reticulum Ca2+ isoform

SVCT:

sodium-dependent vitamin C transporters

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Authors and Affiliations

  1. Department of Biology, McMaster University, Hamilton

    Kelly A. Best, Melanie E. Holmes & Ashok K. Grover

  2. Department of Medicine, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5

    Sue E. Samson, James Mwanjewe & Ashok K. Grover

  3. Department of Physiology, University of Western Ontario, London, Ontario, Canada

    John X. Wilson & S. Jeffrey Dixon

Authors
  1. Kelly A. Best
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  2. Melanie E. Holmes
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  3. Sue E. Samson
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  4. James Mwanjewe
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  5. John X. Wilson
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  6. S. Jeffrey Dixon
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  7. Ashok K. Grover
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Correspondence to Ashok K. Grover.

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Best, K.A., Holmes, M.E., Samson, S.E. et al. Ascorbate uptake in pig coronary artery endothelial cells. Mol Cell Biochem 271, 43–49 (2005). https://doi.org/10.1007/s11010-005-3442-0

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  • DOI: https://doi.org/10.1007/s11010-005-3442-0

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