Bioluminescent Metabolite Detection Assays for Studies of Liver Function
Part # PS473
Abstract
The liver plays a key role in the metabolism of both lipids and carbohydrates. It maintains blood glucose levels by storing glucose when levels are high (glycogenesis) or releasing glucose when levels are low (glycogenolysis and gluconeogenesis). The liver is also the central hub for the uptake, formation, and export of lipids in the form of lipoproteins, and the target of aberrant lipid accumulation in nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH).
To monitor aspects of these functions and pathologies we developed bioluminescent glucose, glucose uptake, and triglyceride assays. The assays share a core reaction sequence that couples luciferase pro-substrate activation to light output that is read on a luminescence plate reader. In the presence of the metabolite of interest, NAD(P), and luciferase, a metabolite selective dehydrogenase reduces the prosubstrate to form NAD(P)H and a luciferin substrate that drives luciferase activity.