Simultaneous Analysis of Major Coenzymes of Cellular Redox Reactions and Energy Using ex Vivo 1H NMR Spectroscopy

Coenzymes of cellular redox reactions and cellular energy mediate biochemical reactions fundamental to the functioning of all living cells. Despite their immense interest, no simple method exists to gain insights into their cellular concentrations in a single step.

Major Coenzymes of Cellular Redox Reactions and Energy Using ex Vivo 1H NMR 1We show that a simple 1H NMR experiment can simultaneously measure oxidized and reduced forms of nicotinamide adenine dinucleotide (NAD+ and NADH), oxidized and reduced forms of nicotinamide adenine dinucleotide phosphate (NADP+ and NADPH), and adenosine triphosphate (ATP) and its precursors, adenosine diphosphate (ADP) and adenosine monophosphate (AMP), using mouse heart, kidney, brain, liver, and skeletal muscle tissue extracts as examples.

Major Coenzymes of Cellular Redox Reactions and Energy Using ex Vivo 1H NMR 2

The analysis protocol along with the annotated characteristic fingerprints for each coenzyme is provided for easy identification and absolute quantification using a single internal reference for routine use. The ability to visualize the ubiquitous coenzymes fundamental to cellular functions, simultaneously and reliably, offers a new avenue to interrogate the mechanistic details of cellular function in health and disease.


Citation & Full Text

Nagana Gowda, G.A., Abell, L., Lee, C.F., Tian, R., Raftery, D.: Simultaneous Analysis of Major Coenzymes of Cellular Redox Reactions and Energy Using Ex Vivo 1H NMR Spectroscopy, Anal. Chem. 88:4817-24, 2016. PMCID: PMC4857157