Abstract

Monitoring Cellular Nucleotide Levels and its Relevance to Cellular Metabolic Function using Bioluminescence-based Strategies

Said Goueli, Clinical Professor & Research Fellow, Promega Corporation

Adenine nucleotides are major determinants of the energy status of the cell and thus any modulation of their cellular concentration has significant consequences to cellular metabolism, cellular growth and cell death. ATP generating enzymes are usually involved in anabolic processes while ATP consuming enzymes are involved in catabolic processes. Monitoring these metabolites in cellular settings has been solely based on HPLC which is of limited throughput and it is time consuming. We have developed a homogenous and bioluminescent strategy to monitor these cellular metabolites using robust HTS formatted and easy to use protocol. We have demonstrated the application of this technology in monitoring the cellular levels of AMP, ADP, and ATP under various physiological conditions and in several cell lines. We will also show the effect of altered cellular metabolites on cell viability and other enzymatic activities that are susceptible to changes in cellular adenine nucleotides.