Methods for measuring CoA and CoA derivatives in biological samples

Abstract

CoA (coenzyme A) is a ubiquitous and essential cofactor that acts as an acyl group carrier in biochemical reactions. Apart from participating in numerous metabolic pathways as substrates and intermediates, CoA and a number of its thioester derivatives, such as acetyl-CoA, can also directly regulate the activity of proteins by allosteric mechanisms and by affecting protein acetylation reactions. Cellular levels of CoA and CoA thioesters change under various physiological and pathological conditions. Defective CoA biosynthesis is implicated in NBIA (neurodegeneration with brain iron accumulation). However, the exact role of CoA in the pathogenesis of NBIA is not well understood. Accurate and reliable assays for measuring CoA species in biological samples are essential for studying the roles of CoA and CoA derivatives in health and disease. The present mini-review discusses methods that are commonly used to measure CoA species in biological samples.

Metabolic regulation of gene expression through histone acylations

“…Methods for the purification, stabilization and measurement of acyl-CoA species have been developed 104 and successfully utilized in cell cultures to study the metabolic regulation of histone modifications 12,36,105,106 . Applying these techniques to a broader range of physiological states should provide valuable information about the dynamic range of the lesser-studied acyl-CoAs.…”

Coenzyme A and protein CoAlation levels are regulated in response to oxidative stress and during morphogenesis in Dictyostelium discoideum

“…CoA extracts were re-suspended in 1 mM DTT and centrifuged for 5 min at 14 000 rpm and 20 °C. CoA level was measured by a modification of the recycling assay developed by Allred and Guy [20,21]. Sets of CoA standards (0.03 μM, 0.1 μM, 0.3 μM, 1 μM, 3 μM, 9 μM) were freshly prepared by serial dilutions of a standard stock solution in 10 mM potassium phosphate buffer (pH 7) and 1 mM DTT.…”

Simultaneous quantification of coenzyme A and its salvage pathway intermediates in in vitro and whole cell-sourced samples

“…These values compare very favourably with those obtained for analysis of CoA (the only analyte in this study for which such parameters have been published) by other methods. 10 For example, CoA has been quantied by HPLC following derivatization with SBD-F with a reported LOD of 120 pmol, 22 and with an LOD of 1.3 nmol using an LC/MS-based technique. 23 Only commercially-available ELISA kits claim lower LODs (as low as 18.3 fmol) for the quantication of free CoA.…”

“…9 The quantication of CoA in biological samples has received wide attention as highlighted by a recent review. 10 The most practical of the CoA analysis methods usually involve separation by HPLC followed by UV-or MS-based detection, or uorescence detection following pre-or post-column derivatisation with a uorophore. However, these methods cannot all be applied with equal success to the CoA biosynthetic intermediates, and consequently the analytical performance of these methods has not been critically evaluated with these compounds.…”