Rapid formation of Nε-(carboxymethyl)lysine (CML) from ribose depends on glyoxal production by oxidation†

Abstract

N ^ε-(Carboxymethyl)lysine (CML) is a major advanced glycation end-product (AGE) involved in protein dysfunction and inflammation in vivo. Its accumulation increases with age and is enhanced with the pathogenesis of diabetic complications. Therefore, the pathways involved in CML formation should be elucidated to understand the pathological conditions involved in CML. Ribose is widely used in glycation research because it shows a high reactivity with proteins to form AGEs. We previously demonstrated that ribose generates CML more rapidly than other reducing sugars, such as glucose; however, the underlying mechanism remains unclear. In this study, we focused on the pathway of CML formation from ribose. As a result, glyoxal (GO) was the most abundant product generated from ribose among the tested reducing sugars and was significantly correlated with CML formation from ribose-modified protein. The coefficient of determination (R²) for CML formation between the ribose-modified protein and Amadori products or the ribose degradation product (RDP)-modified protein was higher for the RDP-modified protein. CML formation from ribose degradation products (RDP) incubated with protein significantly correlated with CML formation from GO-modified protein (r_s = 0.95, p = 0.0000000869). GO and CML formation were inhibited by diethylenetriaminepentaacetic acid (DTPA) and enhanced by iron chloride. Additionally, flavonoid compounds such as isoquercetin, which are known to inhibit CML, also inhibited GO formation from ribose and CML formation. In conclusion, ribose undergoes auto-oxidation and oxidative cleavage between C-2 and C-3 to generate GO and enhance CML accumulation.