Rapid determination of tricarboxylic acid cycle enzyme activities in biological samples.

BMC Biochemistry
Sergio GoncalvesP Rustin

Abstract

In the last ten years, deficiencies in tricarboxylic acid cycle (TCAC) enzymes have been shown to cause a wide spectrum of human diseases, including malignancies and neurological and cardiac diseases. A prerequisite to the identification of disease-causing TCAC enzyme deficiencies is the availability of effective enzyme assays. We developed three assays that measure the full set of TCAC enzymes. One assay relies on the sequential addition of reagents to measure succinyl-CoA ligase activity, followed by succinate dehydrogenase, fumarase and, finally, malate dehydrogenase. Another assay measures the activity of alpha-ketoglutarate dehydrogenase followed by aconitase and isocitrate dehydrogenase. The remaining assay measures citrate synthase activity using a standard procedure. We used these assays successfully on extracts of small numbers of human cells displaying various severe or partial TCAC deficiencies and on frozen heart homogenates from heterozygous mice harboring an SDHB gene deletion. This set of assays is rapid and simple to use and can immediately detect even partial defects, as the activity of each enzyme can be readily compared with one or more other activities measured in the same sample.

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Citations

Mar 11, 2011·World Journal of Gastroenterology : WJG·Jun-Duo HuJin-Lian Chen
May 3, 2014·Systems Biology in Reproductive Medicine·Min TangChang-Jun Yin
Dec 3, 2014·Frontiers in Cell and Developmental Biology·Ryan J Mailloux, William G Willmore
Nov 2, 2013·The Journal of Biological Chemistry·Valérie Desquiret-DumasVincent Procaccio
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Feb 26, 2016·American Journal of Physiology. Endocrinology and Metabolism·Edward OjukaPrisca Ofure Osiki
Sep 6, 2020·Nature Communications·Elizabeth D LuczakMark E Anderson
Mar 23, 2018·Neurology. Genetics·Cecilia MarelliClaire Guissart
Oct 28, 2019·Molecular Genetics and Metabolism·Parith WongkittichoteKimberly A Chapman
Nov 25, 2020·Free Radical Biology & Medicine·Pierre MounkoroBrigitte Meunier

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BETA
biopsies

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