Development of A Sensitive Biochemical Assay for Quantitation of Methylglyoxal
Location
Village Square
Department
Chemistry
Abstract
Methylglyoxal (MG), is one of the naturally existing reactive carbonyl species (RCSs) generated through various metabolic, enzymatic and oxidative reactions in living systems. It reacts with various endogenous nucleophiles such as proteins and nucleic acids, and form advanced glycation end products (AGEs) that leads to cellular damage and apoptosis at various organs. Metabolism of glucose, termed glycolysis, is one of the major pathways of MG production. Elevated MG have been found in patients with diabetes mellitus, which is characterized by high levels of blood glucose leading to upregulated glycolysis. The elevated level of MG and other RCSs are associated with many other diseases including cardiovascular problems, kidney dysfunctions and chronic inflammation. In order to study MG’s role in these biochemical and cellular pathways, an analysis method that can quantify MG concentrations in various environments is in high demand. In this work we aim to develop a sensitive yet simple, cost-effective and user-friendly method to analyze MG levels in various solution samples including tissue culture media and biofluids. A chemical probe, 1,8-diaminonaphthalene (DAN), was shown to react with MG and generate a highly fluorescent product that can be detected at emission wavelength 420nm when excited at wavelength 335nm. The reaction is fast and demonstrates a good linearity between the MG concentrations and fluorescence intensities measured. This method can be easily tailored to monitor the level of MG during MG’s interactions with various components in a chemical or biological sample. We will demonstrate its use in quantifying MG level change in mammalian cell culture.
Faculty Sponsor
Jing Su, Northeastern Illinois University
Development of A Sensitive Biochemical Assay for Quantitation of Methylglyoxal
Village Square
Methylglyoxal (MG), is one of the naturally existing reactive carbonyl species (RCSs) generated through various metabolic, enzymatic and oxidative reactions in living systems. It reacts with various endogenous nucleophiles such as proteins and nucleic acids, and form advanced glycation end products (AGEs) that leads to cellular damage and apoptosis at various organs. Metabolism of glucose, termed glycolysis, is one of the major pathways of MG production. Elevated MG have been found in patients with diabetes mellitus, which is characterized by high levels of blood glucose leading to upregulated glycolysis. The elevated level of MG and other RCSs are associated with many other diseases including cardiovascular problems, kidney dysfunctions and chronic inflammation. In order to study MG’s role in these biochemical and cellular pathways, an analysis method that can quantify MG concentrations in various environments is in high demand. In this work we aim to develop a sensitive yet simple, cost-effective and user-friendly method to analyze MG levels in various solution samples including tissue culture media and biofluids. A chemical probe, 1,8-diaminonaphthalene (DAN), was shown to react with MG and generate a highly fluorescent product that can be detected at emission wavelength 420nm when excited at wavelength 335nm. The reaction is fast and demonstrates a good linearity between the MG concentrations and fluorescence intensities measured. This method can be easily tailored to monitor the level of MG during MG’s interactions with various components in a chemical or biological sample. We will demonstrate its use in quantifying MG level change in mammalian cell culture.