Purification and properties of retinol- and retinoic acid-binding proteins from a transplantable mouse colon tumor.
Cellular retinol-binding protein and retinoic acid-binding protein, the possible mediators of the action of retinoids in epithelial differentiation and control of tumorigenesis, have been reproducibly purified from mouse colon tumor 26, and some of their properties were studied. The main steps of purification involved acid-precipitation, DEAE-Sephadex, CM-cellulose and Sephadex G-100 chromatography. About 2 mg of the binding proteins were isolated from 60 g tumor. The purified preparations showed only two protein bands on polyacrylamide gel electrophoresis. The two binding proteins were partially resolved by sedimentation equilibrium technique; but was completely separable by preparative electrophoresis in the presence of sodium dodecyl sulfate. The retinol- and retinoic acid-binding proteins are presumably monomers with molecular weights of 15,500 and 14,600, respectively, as determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. On gel filtration however, both the binding proteins retarded to the same molecular size of 17,800. On preparative columns, both the proteins expressed the same isoelectric pH, 4.5. Both proteins of the tumor possessed functional thiol groups. The mercurial inhibition of the binding capacity of the proteins for their ligands was reversible upon treatment with thiol compounds.
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification*; D002499:Centrifugation, Density Gradient; D003110:Colonic Neoplasms / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D051379:Mice; D008970:Molecular Weight; D018168:Receptors, Retinoic Acid; D012177:Retinol-Binding Proteins / Q000302:isolation & purification*; D054840:Retinol-Binding Proteins, Cellular; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014212:Tretinoin / Q000302:isolation & purification*
D002352:Carrier Proteins; D018168:Receptors, Retinoic Acid; D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D013438:Sulfhydryl Compounds; D012967:Sodium Dodecyl Sulfate; D014212:Tretinoin