Study of formation of advanced glycation endproducts by the reaction between guanine tri-phosphate and reducing sugars
The Advanced Glycation Endproducts (AGE’s) are formed between the interaction of amino groups in proteins and amino acids with keto group of a sugar under normal physiological conditions. These AGE products are believed to be responsible for the numerous complications in humans observed in diabetes, Alzheimer disease, cataract formation, ischemic heart attack and aging. This thesis focuses on monitoring the formation of the AGE products formed by Low Molecular weight DNA neucleobases with the reducing sugars, Methyl Glyoxal and Glucosamine. DNA nucleotides such as GuanosineTriPhosphate (GTP) are used as the model nucleotide substrates reacting with Methyl Glyoxal. Samples were prepared at concentrations of 5 mM, 10mM and 20mM of Methyl Glyoxal (MG) with 5mM of GTP. Samples of 1:1 mixture of MG/GlcN and GTP were incubated and collected at various times. Methods were developed for UV, Fluorescence, and HPLC for each of these techniques using various scouting experiments. Ammonium Acetate buffer is used for preparation of samples and reaction mixtures between MG/GlcN and GTP. There were no clear trends obtained during UV and Fluorescence analysis. Further the HPLC results show only one major peak. After significant analysis, it was concluded that there is strong interference between Ammonium acetate buffer and GTP/MG and GlcN. Due to chemical reaction, the amount of AGEs formed are very minimal. Only some signs of AGE formation is obtained at higher concentration at day 21 samples of GTP/GlcN mixture. To remedy the interference of ammonium buffer, experiments should be performed using PBS buffer instead of Ammonium acetate buffer.
"Study of formation of advanced glycation endproducts by the reaction between guanine tri-phosphate and reducing sugars"
Dissertations and Master's Theses (Campus Access).