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Ascorbic acid transport and distribution in human B lymphocytes

P Bergsten1, R Yu, J Kehrl

  • 1Laboratory of Cell Biology and Genetics, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.

Archives of Biochemistry and Biophysics
|February 20, 1995
PubMed
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Human B lymphocytes transport ascorbic acid (vitamin C) via a high-affinity system. This vitamin C transport mechanism is efficient at normal human plasma concentrations.

Area of Science:

  • Cellular biology
  • Biochemistry
  • Immunology

Background:

  • Ascorbic acid (vitamin C) is essential for various cellular functions.
  • Understanding vitamin C transport in immune cells like B lymphocytes is crucial for immune health.
  • Previous research has not fully elucidated the mechanisms of vitamin C uptake in human B lymphocytes.

Purpose of the Study:

  • To investigate the transport kinetics and characteristics of ascorbic acid in human B lymphocytes.
  • To determine the subcellular localization of intracellular ascorbic acid within these cells.
  • To assess the physiological relevance of the identified transport mechanisms.

Main Methods:

  • Utilized radiolabeled ascorbic acid to measure transport rates in isolated human B lymphocytes.

Related Experiment Videos

  • Performed kinetic analyses to determine Michaelis-Menten parameters (Km, Vmax).
  • Investigated the effects of inhibitors (e.g., carbonylcyanide-p-trifluoromethoxyphenylhydrazone, ouabain) and temperature on transport.
  • Conducted subcellular fractionation to determine the localization of ascorbic acid.

    • Main Results:

    • Identified a high-affinity transport system for ascorbic acid in human B lymphocytes with specific kinetic parameters (Km: 7-10 microM, Vmax: 0.14 mM/h).
    • Demonstrated that this transport is saturable, concentration-dependent, temperature-dependent, and inhibited by specific compounds, indicating an active process.
    • Observed that >90% of intracellular ascorbic acid is localized in the cytosol, not protein-bound.
    • Found that the high-affinity transport kinetics are suitable for physiological plasma concentrations of vitamin C.

    Conclusions:

    • Human B lymphocytes possess a high-affinity, active transport system for ascorbic acid.
    • This transport system is capable of accumulating vitamin C against a concentration gradient.
    • The identified transport mechanism is physiologically relevant for maintaining adequate intracellular vitamin C levels in B lymphocytes.