Characterization and purification of a mammalian osmoregulatory protein, aldose reductase, induced in renal medullary cells by high extracellular NaCl.
GRB-PAP1 is a continuous line of epithelial cells derived from a rabbit renal inner medulla. Elevation of the NaCl concentration in the medium bathing these cells strongly induced the expression of a soluble protein with an apparent molecular mass of 39 kDa. The protein, purified by affinity chromatography with Amicon Matrex Gel Orange A, had enzyme activity characteristic of aldose reductase (alditol:NADPH+ oxidoreductase, EC 1.1.1.21). Goat antiserum against this purified aldose reductase selected the 39-kDa band from immunoblots of cells grown in a medium containing high NaCl. When the osmolality of the medium was increased by adding NaCl, the amount of aldose reductase protein and the aldose reductase activity increased together from very low to sustained high levels over several days. The aldose reductase protein was more than 10% of the soluble cell protein when cells were propagated in medium made hyperosmotic by adding NaCl to increase medium osmolality to 600 mosm.kg-1.
D000449:Aldehyde Reductase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004790:Enzyme Induction; D004848:Epithelium / Q000201:enzymology; D005779:Immunodiffusion; D007679:Kidney Medulla / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D008970:Molecular Weight; D000073718:Pancreatitis-Associated Proteins; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology*; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism*
D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; D012965:Sodium Chloride; D013401:Sugar Alcohol Dehydrogenases; D000449:Aldehyde Reductase