Detection and determination of glyceraldehyde-derived advanced glycation end product.
Protein is modified by carbonyl compound in the Maillard reaction, and the irreversible structure is formed as the advanced glycation end product (AGE). We identified GLAP (glyceraldehyde-derived pyridinium compound) as an AGE formed from glyceraldehyde and lysine residue of protein. In the present study, we investigated detection and determination of GLAP from glycated protein using fluorescence HPLC method. Albumin (BSA) and carbonyls (glyceraldehyde, glycolaldehyde, methylglyoxal, glyoxal, three pentoses or three hexoses) were dissolved in phosphate buffed solution (pH 7.4), and incubated at 37 degrees C for a week. GLAP was formed only in the glyceraldehyde-modified BSA. It is suggested that GLAP was specific AGE derived from glyceraldehyde. In addition, GLAP depressed the intracellular glutathione level and induced the reactive oxygen species (ROS) in HL-60 cells. GLAP caused the oxidative stress. Therefore, GLAP will be a biomarker in the AGE related disease such as diabetic complications or chronic renal failure.
Usui
Teruyuki
T
Department of Agricultural Chemistry, Faculty of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
Shimohira
Kazuhito
K
Watanabe
Hirohito
H
Hayase
Fumitaka
F
D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D017127:Glycation End Products, Advanced / Q000032:analysis* / Q000378:metabolism*; D005985:Glyceraldehyde / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D008239:Lysine; D018384:Oxidative Stress; D012710:Serum Albumin, Bovine / Q000737:chemistry
D017127:Glycation End Products, Advanced; D012710:Serum Albumin, Bovine; D005985:Glyceraldehyde; D008239:Lysine