TY - JOUR
T1 - Purification and properties of aldose reductase and aldehyde reductase from EHS tumor cells
AU - Tanimoto, Tsuyoshi
AU - Sato, Sanai
AU - Kador, Peter F.
PY - 1990/2/1
Y1 - 1990/2/1
N2 - Engelbreth-Holm-Swarm (EHS) tumor cells were utilized as a model for investigating the production of basement membrane components. These cells contain two immunologically distinct NADPH-dependent reductases, aldose reductase (EC 1.1.1.21) and aldehyde reductase (EC 1.1.1.2), which were purified to apparent homogeneity by a combination of procedures which included ammonium sulfate fractionation, Sephadex G-75 gel filtration, Matrex Gel Orange A affinity chromatography, and chromatofocusing on Phannacia Mono P. The molecular weights of aldose and aldehyde reductases were estimated to be 38K and 40K, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Substrate specificity studies showed that both enzymes were capable of reducing a variety of aldehydes to their respective alcohols; however, only aldehyde reductase oxidized l-gulonic acid. Surprisingly, both enzymes showed similar reactivities with d-glucose and d-galactose, suggesting that both aldose and aldehyde reductases may contribute to sorbitol production in the EHS tumor cell. The activities of both enzymes were increased by the presence of sulfate ion, but chloride ion decreased the activity of aldose reductase. Both aldose and aldehyde reductases were inhibited by a series of structurally diverse aldose reductase inhibitors.
AB - Engelbreth-Holm-Swarm (EHS) tumor cells were utilized as a model for investigating the production of basement membrane components. These cells contain two immunologically distinct NADPH-dependent reductases, aldose reductase (EC 1.1.1.21) and aldehyde reductase (EC 1.1.1.2), which were purified to apparent homogeneity by a combination of procedures which included ammonium sulfate fractionation, Sephadex G-75 gel filtration, Matrex Gel Orange A affinity chromatography, and chromatofocusing on Phannacia Mono P. The molecular weights of aldose and aldehyde reductases were estimated to be 38K and 40K, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Substrate specificity studies showed that both enzymes were capable of reducing a variety of aldehydes to their respective alcohols; however, only aldehyde reductase oxidized l-gulonic acid. Surprisingly, both enzymes showed similar reactivities with d-glucose and d-galactose, suggesting that both aldose and aldehyde reductases may contribute to sorbitol production in the EHS tumor cell. The activities of both enzymes were increased by the presence of sulfate ion, but chloride ion decreased the activity of aldose reductase. Both aldose and aldehyde reductases were inhibited by a series of structurally diverse aldose reductase inhibitors.
UR - http://www.scopus.com/inward/record.url?scp=0025129179&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0025129179&partnerID=8YFLogxK
U2 - 10.1016/0006-2952(90)90049-Q
DO - 10.1016/0006-2952(90)90049-Q
M3 - Article
C2 - 2106320
AN - SCOPUS:0025129179
SN - 0006-2952
VL - 39
SP - 445
EP - 453
JO - Biochemical Pharmacology
JF - Biochemical Pharmacology
IS - 3
ER -