Rapid characterization of cellular retinoid-binding proteins by high-performance size-exclusion chromatography.
A high-performance size-exclusion chromatographic method was developed for the analysis of cellular retinol and retinoic acid-binding proteins. The chromatographic analysis of the retinol-retinol-binding protein complex or the retinoic acid-retinoic acid-binding protein complex requires 15 min. The use of high-specific-radioactivity retinoid ligand (30-40 Ci/mmol) allows routine detection of 25 fmol of retinoid-binding protein/mg of cytosolic protein. Thus, this method provides a rapid alternative to sucrose density gradient sedimentation analysis of the cellular retinoid-binding proteins. High-performance size-exclusion chromatography is well suited to screening novel tissues, tumors, and cell lines for the presence of retinoid-binding proteins and to the further characterization of these cellular proteins. This method was applied to the characterization of cellular retinol- and retinoic acid-binding proteins in fetal rabbit tissues. Both retinol- and retinoic acid-binding proteins were detected in fetal rabbit brain, intestine, kidney, and lung at a gestational age of 28 days. Neither retinoid-binding protein was detected in 28-day-old fetal rabbit placenta. Specific retinoic acid binding in fetal intestinal cytosol decreased as a function of increasing gestational age.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003600:Cytosol; D005260:Female; D005333:Fetus; D005865:Gestational Age; D007421:Intestine, Small / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D008297:Male; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D018168:Receptors, Retinoic Acid; D012176:Retinoids / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D014018:Tissue Distribution; D014212:Tretinoin / Q000378:metabolism
D002352:Carrier Proteins; D018168:Receptors, Retinoic Acid; D012176:Retinoids; D014212:Tretinoin