Involvement of a receptor-mediated component in cellular translocation of riboflavin.

This study addresses the transport mechanism of riboflavin (vitamin B(2)) 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 (K(m): 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.

• Publication year

  2000

• Venue

  Journal of Pharmacology and Experimental Therapeutics

• Publication date

  2000-07-01

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0022-3565(24)39046-9PMID 10871302
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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  1999cited by this paper
• The biochemical and physiological characteristics of receptors.

  1998cited by this paper
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• Folate-mediated targeting of therapeutic and imaging agents to cancers.

  1998cited by this paper
• Endocytosis and transcytosis.

  1998cited by this paper
• Microtubule-dependent vesicle transport: modulation of channel and transporter activity in liver and kidney.

  1998influential reference
• Binding and Transcytosis of Botulinum Neurotoxin by Polarized Human Colon Carcinoma Cells*

  1998cited by this paper
• A human colonic cell line sharing similarities with enterocytes as a model to examine oral absorption: advantages and limitations of the Caco-2 model.

  1997cited by this paper
• Endocytosis.

  1997cited by this paper
• Measurements and characteristics of intestinal riboflavin transport.

  1997cited by this paper
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• Pharmacokinetics of orally and intravenously administered riboflavin in healthy humans.

  1996influential reference
• Cultured intestinal epithelial cell models.

  1996influential reference
• Use of the intestinal and hepatic bile acid transporters for drug delivery

  1996cited by this paper
• Folate receptors.

  1996cited by this paper
• Riboflavin-Enhanced Transport of Serum Albumin Across the Distal Pulmonary Epithelium

  1996cited by this paper
• Membrane protein trafficking through the common apical endosome compartment of polarized Caco-2 cells.

  1995cited by this paper
• Riboflavin uptake by native Xenopus laevis oocytes.

  1995influential reference
• Mammalian ion‐coupled solute transporters.

  1995cited by this paper
• Determination of Transport Rates for Arginine and Acetaminophen in Rabbit Intestinal Tissues in Vitro

  1994cited by this paper
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  1994cited by this paper
• Mechanism of riboflavine uptake by Caco-2 human intestinal epithelial cells.

  1994cited by this paper
• Uptake of riboflavin by intestinal basolateral membrane vesicles: a specialized carrier-mediated process.

  1993cited by this paper
• Uptake of riboflavin across the brush border membrane of rat intestine: regulation by dietary vitamin levels.

  1993cited by this paper
• Biotin transport in a human intestinal epithelial cell line (Caco-2).

  1993influential reference
• Brefeldin A enhances receptor-mediated transcytosis of transferrin in filter-grown Madin-Darby canine kidney cells.

  1992cited by this paper
• Effects of brefeldin A on endocytosis, transcytosis and transport to the Golgi complex in polarized MDCK cells

  1992cited by this paper
• Transport of riboflavin in human intestinal brush border membrane vesicles.

  1991cited by this paper
• Uptake of riboflavin by rat intestinal mucosa in vitro.

  1990cited by this paper
• Transport of bile acids in a human intestinal epithelial cell line, Caco-2.

  1990cited by this paper
• The transport of vitamin B12 through polarized monolayers of Caco-2 cells.

  1990influential reference
• A specific sorting signal is not required for the polarized secretion of newly synthesized proteins from cultured intestinal epithelial cells

  1988cited by this paper
• Intestinal absorption of riboflavin, studied by an in situ circulation system using radioactive analogues.

  1988cited by this paper
• Chicken riboflavin-binding protein. cDNA sequence and homology with milk folate-binding protein.

  1988cited by this paper
• The physiology of transferrin and transferrin receptors.

  1987cited by this paper
• Receptor-mediated folate accumulation is regulated by the cellular folate content.

  1986cited by this paper
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  1982cited by this paper

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