Acetyl-Coenzyme A UV-Spectrophotometric Assay

Add time:08/24/2019    Source:sciencedirect.com

Publisher SummaryThis chapter provides an overview of acetyl-CoA. It is the common intermediate in the oxidation of carbohydrates, fatty acids, and certain amino acids, and it is the point of entry for terminal oxidation in the citric acid cycle. Acetyl-CoA is also the precursor for the biosynthesis of fatty acids, acetoacetate, and steroids. Consequently, it is found in all living cells. It has not been detected in extracellular fluids (blood). Assay methods for acetyl-CoA are based on the enzymatic acetylation of aromatic amines, on the arsenolysis of acetyl-CoA catalyzed by phosphotransacetylase (PTA), or on the synthesis of citrate from oxaloacetate in the presence of citrate synthase (CS). Only a small portion of the total CoA content of tissues is in the form of acetyl-CoA; spectrophotometric assays, therefore, require relatively large amounts of sample. Acetoaeetyl-CoA occurs as an intermediate in the β-oxidation of fatty acids and in the synthesis of 3-hydroxy-3-methylglutaryl-CoA in cell-free systems. Acrylyl-CoA is aminated with NH4C1 and acrylyl-CoA aminase with a corresponding decrease in extinction at 263 nm.

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