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.
National Eye Institute, Laboratory of Mechanism of Ocular Disease, National Institutes of Health, Bethesda, Maryland 20892.
K
K
Karino
P F
PF
Kador
B
B
Berkowitz
R S
RS
Balaban
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