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 chromat...

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Bibliographic Details
Published in:The Journal of biological chemistry 1987-10, Vol.262 (29), p.14255-14259
Main Authors: Bedford, J J, Bagnasco, S M, Kador, P F, Harris, H W, Burg, M B
Format: Article
Language:English
Subjects:
Aldehyde Reductase - biosynthesis
Aldehyde Reductase - isolation & purification
Aldehyde Reductase - metabolism
aldose dehydrogenase
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Cell Line
Enzyme Induction
Enzymes and enzyme inhibitors
Epithelium - enzymology
Fundamental and applied biological sciences. Psychology
Immunodiffusion
Kidney Medulla - drug effects
Kidney Medulla - enzymology
Kinetics
medulla
Molecular Weight
osmoregulation
Oxidoreductases
Pancreatitis-Associated Proteins
Rabbits
Sodium Chloride - pharmacology
Substrate Specificity
Sugar Alcohol Dehydrogenases - metabolism
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)47931-8