Abstract
The non-enzymatic reaction between reducing sugars and proteins has received increased attention in nutritional and medical research recently. In the current manuscript, effect of glycation in structural changes of human serum albumin (HSA) by the metabolites of glucose such as glyoxal, methylglyoxal and glyceraldehyde was studied using different spectroscopy techniques. Glycation of HSA was monitored by following advanced glycation end-products (AGEs) fluorescence changes, HSA intrinsic fluorescence measurement, extrinsic fluorescence using 8-analino 1-nephthlene sulfonic acid (ANS) dye, and circular dichroism (CD) studies. AGEs were formed within 7 days of incubation with glyoxal, methylglyoxal and glyceraldehyde. However, methylglyoxal induced significant structural changes in HSA compared with glyoxal and glyceraldehydes. Moreover, ANS binding to native and glycated-HSA showed difference in binding pattern of these metabolites to HSA. The CD spectrum revealed changes in the secondary structure of HSA upon glycation when compared to native HSA. Furthermore, the MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay established the cytotoxicity of the glycated- HSA towards human liver carcinoma (HepG2) cell lines via the production of reactive oxygen species.