The presence of 2-keto-3-deoxygluconic acid and oxoaldehyde dehydrogenase activity in human erythrocytes.
2-Keto-3-deoxygluconic acid (3-DGA) is produced from 3-deoxyglucosone (3-DG:a highly reactive glycation intermediate) through oxidation by the enzyme oxoaldehyde dehydrogenase (OAD) in animals. We developed a specific assay method for 3-DGA using high-performance liquid chromatography [Fujii, E. et al. (1994) J. Chromatogr. B 660, 265-270] and measured it in the hemolysate and plasma of diabetic patients and healthy subjects. Both human erythrocytes and plasma contained considerable amounts of 3-DGA. However, human erythrocyte contained about 30-50 times higher 3-DGA than human plasma did and also had the same ability to convert 3-DG to 3-DGA as OAD had. Erythrocyte 3-DGA levels of diabetic patients were 990 +/- 370 nmol/gHb (n = 57, Mean +/- SD) and were significantly higher compared with healthy subjects (527 +/- 194 nmol/gHb, n = 7, p < 0.01). In all diabetic patients and healthy subjects (n = 64), there was only one patient who had a very low level of erythrocyte 3-DGA and lacked the ability to convert 3-DG to 3-DGA. When erythrocytes were incubated at 37 degrees C for 8 hours in phosphate buffer containing 0.35 mM 3-DG, 3-DG was easily taken into the erythrocytes and was converted to 3-DGA. Our results suggest the contribution of OAD not only to the prevention of glycation of hemoglobin but also to that of blood vessels by scavenging plasma 3-DG into erythrocytes.
Fujii
E
E
Department of Internal Medicine, School of Medicine, Kitasato University, Kanagawa, Japan.
Iwase
H
H
Ishii-Karakasa
I
I
Yajima
Y
Y
Hotta
K
K
D000328:Adult; D000368:Aged; D000429:Alcohol Oxidoreductases / Q000097:blood*; D001692:Biological Transport; D002851:Chromatography, High Pressure Liquid; D003922:Diabetes Mellitus, Type 1 / Q000097:blood* / Q000201:enzymology; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000201:enzymology; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D005942:Gluconates / Q000097:blood*; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D012016:Reference Values
D005942:Gluconates; C002957:2-keto-3-deoxygluconate; D000429:Alcohol Oxidoreductases; C019642:alcohol dehydrogenase (NADP+)