19F NMR quantitation of lens aldose reductase activity using 3-deoxy-3-fluoro-D-glucose.
In the present study we have determined the kinetics of 3-deoxy-3-fluoro-D-glucose (3-FG) as a substrate for the aldose reductase reaction in vitro. In addition, we compared the 3-deoxy-3-fluoro-sorbitol (3-FS) production rates from 3-FG in the intact lens using 19F NMR with conventional aldose reductase determinations in extracts from the same lenses. The affinity of in vitro aldose reductase for 3-FG was approximately 20 times greater (9.3 mM) than that for glucose (188 mM). An excellent correlation between the rate of 3-FS production in the intact canine lens, determined with 19F NMR, and extracted aldose reductase activity was observed. The relatively high affinity of aldose reductase for 3-FG and the correlation of 3-FS production with enzyme activity in the intact lens suggests that 3-FS production from 3-FG detected by 19F NMR could provide an accurate noninvasive determination of aldose reductase activity in the eye lens.
Karino
K
K
National Eye Institute, Laboratory of Mechanism of Ocular Disease, National Institutes of Health, Bethesda, Maryland 20892.
Kador
P F
PF
Berkowitz
B
B
Balaban
R S
RS
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D003847:Deoxyglucose / Q000031:analogs & derivatives / Q000378:metabolism; D004285:Dogs; D066298:In Vitro Techniques; D007700:Kinetics; D007908:Lens, Crystalline / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D011799:Quinazolines / Q000494:pharmacology; D013012:Sorbitol / Q000378:metabolism
D011799:Quinazolines; C064920:FR 74366; C007253:3-deoxy-3-fluoro-D-glucose; D013012:Sorbitol; D003847:Deoxyglucose; D000449:Aldehyde Reductase