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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Involvement of a Receptor-Mediated Component in Cellular Translocation of Riboflavin

Se-Ne Huang and Peter W. Swaan
Journal of Pharmacology and Experimental Therapeutics July 2000, 294 (1) 117-125;
Se-Ne Huang
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Peter W. Swaan
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Abstract

This study addresses the transport mechanism of riboflavin (vitamin B2) across intestinal epithelium in the presence and absence of pharmacologically active compounds. A polarized transport process with a 6-fold higher basolateral (BL)-to-apical (AP) flux was observed in both a human intestinal cell model (Caco-2) and rat intestinal tissue. Riboflavin-specific translocation systems on both the AP and BL cell surfaces were saturable with affinity values close to most receptors (Km: 9.72 ± 0.85 and 4.06 ± 0.03 nM, respectively). Pharmacological agents known to alter intracellular endocytic events were used to examine the potential involvement of receptor-mediated events. Nocodazole significantly inhibited AP uptake (58.4%), BL-to-AP riboflavin (56.7%) and fluorescein isothiocyanate-labeled transferrin (FITC-Tf) (31.8%) transport without affecting mannitol or cholic acid transport, whereas AP-to-BL riboflavin (252.8%) and FITC-Tf (145.1%) transport was increased. Brefeldin A significantly enhanced AP-to-BL riboflavin (37.1%) and bidirectional FITC-Tf transport (AP-to-BL: 13-fold; BL-to-AP: 5-fold). without affecting BL-to-AP riboflavin transport. Combined, these data suggest an essential role of microtubule-dependent movement and vesicular sorting component(s) in the bidirectional transport of riboflavin. Dissociation of riboflavin from the cell surface was pH-dependent with significantly higher substrate release at acidic pH, indicating the presence of riboflavin-specific cell surface receptors. In summary, our studies provide biochemical evidence of the involvement of a receptor-mediated mechanism in the cellular translocation of riboflavin.

Footnotes

  • Send reprint requests to: Dr. Peter W. Swaan, Division of Pharmaceutics, College of Pharmacy, 500 West 12th Ave., Columbus OH 43210-1291. E-mail: swaan.1{at}osu.edu

  • Abbreviations:
    RME
    receptor-mediated endocytosis
    FITC-Tf
    fluorescein isothiocyanate-labeled transferrin
    TfR
    transferrin receptor
    AP
    apical
    BL
    basolateral
    BFA
    brefeldin A
    Is.c.
    short-circuit current
    • Received December 16, 1999.
    • Accepted March 10, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 294 (1)
Journal of Pharmacology and Experimental Therapeutics
Vol. 294, Issue 1
1 Jul 2000
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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Involvement of a Receptor-Mediated Component in Cellular Translocation of Riboflavin

Se-Ne Huang and Peter W. Swaan
Journal of Pharmacology and Experimental Therapeutics July 1, 2000, 294 (1) 117-125;

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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Involvement of a Receptor-Mediated Component in Cellular Translocation of Riboflavin

Se-Ne Huang and Peter W. Swaan
Journal of Pharmacology and Experimental Therapeutics July 1, 2000, 294 (1) 117-125;
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