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. (C) 1995 Academic Press, Inc.