Aldose reductase from human skeletal and heart muscle. Interconvertible forms related by thiol-disulfide exchange.
Aldose reductase was purified from human skeletal and heart muscle by a rapid and efficient scheme involving Red Sepharose chromatography, chromatofocusing on Pharmacia PBE 94, and hydroxylapatite high pressure liquid chromatography. The scheme afforded homogeneous enzyme, 65% recovery, in 2 days. All muscle samples express aldose reductase but not the closely related aldehyde reductase. Aldose reductase is isolated in one of two forms that are distinguishable by their kinetic patterns with glyceraldehyde as substrate and which are interconvertible by treatment with dithiothreitol. Both forms are capable of catalyzing the reduction of glucose (Km = 68 mM), and both are highly sensitive to inhibition by aldose reductase inhibitors. The reduction of glucose was shown to be nearly stoichiometric with production of sorbitol (92 +/- 2%). Dialysis of aldose reductase in the absence of thiols or NADP converts it into a form that shows markedly different kinetic properties, including very weak catalytic activity toward glucose and insensitivity to aldose reductase inhibitors. This modified form can be converted back into the native form by dithiothreitol. Thiol titration of the two forms of aldose reductase with Ellman's reagent indicated that two thiol groups were lost when the enzyme was dialyzed in the absence of dithiothreitol or NADP.
Vander Jagt
D L
DL
Department of Biochemistry, University of New Mexico School of Medicine, Albuquerque 87131.
Robinson
B
B
Taylor
K K
KK
Hunsaker
L A
LA
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D006801:Humans; D006882:Hydroxyapatites; D007093:Imidazoles / Q000494:pharmacology; D048289:Imidazolidines; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology*; D009281:Naphthalenes / Q000494:pharmacology; D010793:Phthalazines / Q000494:pharmacology
D006882:Hydroxyapatites; D007093:Imidazoles; D048289:Imidazolidines; D007527:Isoenzymes; D009281:Naphthalenes; D010793:Phthalazines; C040550:tolrestat; C042829:ponalrestat; D017886:Durapatite; D000449:Aldehyde Reductase; C026411:sorbinil