Photoaffinity labeling of retinoic acid-binding proteins.
Retinoid-binding proteins are essential mediators of vitamin A function in vertebrate organisms. They solubilize and stabilize retinoids, and they direct the intercellular and intracellular trafficking, transport, and metabolic function of vitamin A compounds in vision and in growth and development. Although many soluble retinoid-binding proteins and receptors have been purified and extensively characterized, relatively few membrane-associated enzymes and other proteins that interact with retinoids have been isolated and studied, due primarily to their inherent instabilities during purification. In an effort to identify and purify previously uncharacterized retinoid-binding proteins, it is shown that radioactively labeled all-trans-retinoic acid can be used as a photoaffinity labeling reagent to specifically tag two known retinoic acid-binding proteins, cellular retinoic acid-binding protein and albumin, in complex mixtures of cytosolic proteins. Additionally, a number of other soluble and membrane-associated proteins that bind all-trans-[11,12-3H]retinoic acid with high specificity are labeled utilizing the same photoaffinity techniques. Most of these labeled proteins have molecular weights that do not correspond to any known retinoid-binding proteins. Thus, photoaffinity labeling with all-trans-retinoic acid and related photoactivatable retinoids is a method that should prove extremely useful in the identification and purification of novel soluble and membrane-associated retinoid-binding proteins from ocular and nonocular tissues.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
P S
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Bernstein
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Choi
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YC
Ho
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Rando
D000345:Affinity Labels; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry; D003600:Cytosol / Q000737:chemistry; D007526:Isoelectric Point; D008970:Molecular Weight; D009928:Organ Specificity; D010777:Photochemistry; D010857:Pigment Epithelium of Eye / Q000737:chemistry; D018168:Receptors, Retinoic Acid / Q000302:isolation & purification* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014212:Tretinoin / Q000378:metabolism*
D000345:Affinity Labels; D018168:Receptors, Retinoic Acid; D012710:Serum Albumin, Bovine; D014212:Tretinoin