Multi-method quantification of acetyl-CoA and further acyl-CoA species in normal and ischemic rat liver

Tokarska-Schlattner, M.; Zeaiter, N.; Cunin, V.; Attia, S.; Meunier, C.; Kay, L.; Achouri, A.; Hiriart-Bryant, E.; Couturier, K.; Tellier, C.; El Harras, A.; Elena-Herrmann, B.; Khochbin, S.; Le Gouellec, A.; Schlattner, U.

doi:10.1101/2023.09.04.556259

bioRxiv · 10.1101/2023.09.04.556259

Multi-method quantification of acetyl-CoA and further acyl-CoA species in normal and ischemic rat liver

Tokarska-Schlattner, M.·Zeaiter, N.·Cunin, V.·Attia, S.·Meunier, C.·Kay, L.·Achouri, A.·Hiriart-Bryant, E.·Couturier, K.·Tellier, C.·El Harras, A.·Elena-Herrmann, B.·Khochbin, S.·Le Gouellec, A.·Schlattner, U.

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Abstract

Thioesters of coenzyme A (CoA) carrying different acyl chains (acyl-CoAs) are central intermediates of many metabolic pathways and donor molecules for protein lysine acylation. Acyl-CoA species largely differ in cellular concentrations and physico-chemical properties, rendering their analysis challenging. Here we compare several approaches to quantify cellular acyl-CoAs concentrations in normal and ischemic rat liver, using HPLC and LC-MS/MS for multi-acyl-CoA analysis, as well as NMR, fluorimetric and spectrophotometric techniques for quantification of acetyl-CoA. In particular, we describe a simple LC-MS/MS protocol that is suitable for the relative quantification of short and medium chain acyl-CoAs species. We show that ischemia induces specific changes in the short chain acyl-CoA relative concentrations, while mild ischemia (1-2 min), although reducing succinyl-CoA, has little effects on acetyl-CoA, and even increases some acyl-CoAs species upstream of the tricarboxylic acid cycle. In contrast, advanced ischemia (5-6 min) also reduces acetyl-CoA levels. Our approach provides the keys to accessing the acyl-CoA metabolome for a more in-depth analysis of metabolism, protein acylation and epigenetics.

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Tokarska-Schlattner, M., Zeaiter, N., Cunin, V., Attia, S., Meunier, C., Kay, L., Achouri, A., Hiriart-Bryant, E., Couturier, K., Tellier, C., El Harras, A., Elena-Herrmann, B., Khochbin, S., Le Gouellec, A., Schlattner, U.. 2023-09-05. Multi-method quantification of acetyl-CoA and further acyl-CoA species in normal and ischemic rat liver. https://doi.org/10.1101/2023.09.04.556259

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