Identification and quantitation of intracellular retinol and retinoic acid binding proteins in cultured cells

Mark R. Haussler, Carol A. Donaldson, Michael A. Kelly, David J. Mangelsdorf, G. Timothy Bowden, William J. Meinke, Frank L. Meyskens, Neil Sidell

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Although the mechanism whereby vitamin A mediates normal cell differentiation and inhibits tumor cell proliferation is unknown, intracellular receptor-like proteins for retinol and retinoic acid have been implicated in the molecular action of vitamin A. We have assayed these two binding proteins, cellular retinol binding protein (protein R) and cellular retinoic acid binding protein (protein RA), in the cytosolic fraction of various normal and tumor cells via sucrose density gradient centrifugation and saturation analysis. Employing charcoal separation of bound and free tritiated retinoid, the saturation analysis yields an approximate Kd for ligand binding and an estimate of the number of protein R and protein RA molecules per cell. Unique protein R and protein RA macromolecules sedimenting at 2 S with Kd values of 7-42 nM are detected in murine cells (1° epidermal, 3T6 fibroblasts and melanoma) and human neuroblastoma cells. Concentrations of the intracellular binding proteins range from 55 000 to 3 000 000 copies per cell. When one cell line (C-127 mouse mammary) is transformed by bovine papilloma virus, protein RA levels increase from undetectable to 193 000 copies per cell. Assessment of growth inhibition by 10-6 M retinol or retinoic acid in the culture medium reveals that there exists a partial, but not absolute, correlation between the presence of protein R or protein RA and the antiproliferative effect of the particular retinoid in the tested cell lines. We conclude that the 2 S intracellular binding proteins for the retinoids are present in most vitamin A responsive cells, but may not be essential for biologic actions of the vitamin such as growth inhibition in monolayer culture.

Original languageEnglish (US)
Pages (from-to)54-62
Number of pages9
JournalBBA - Molecular Cell Research
Volume803
Issue number1-2
DOIs
StatePublished - Feb 17 1984

Keywords

  • Cellular retinoic acid binding protein
  • Cellular retinol binding protein

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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