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Different inherited enzyme deficiencies in which the oxidation of acyl\u2010CoAs is hampered have been hypothesised to share a phenotype characterised by toxic accumulation of acyl\u2010CoA and a concomitant decline in free CoA (CoASH) levels, whereby CoASH becomes limiting for other metabolic reactions. This is referred to as CoASH sequestration. There is, however, limited experimental evidence for this hypothesis. Using a combination of approaches, we test this hypothesis in medium\u2010chain acyl\u2010CoA dehydrogenase deficiency (MCADD), the most common deficiency of mitochondrial fatty acid oxidation (mFAO), under energetic stress. Both\n                    <jats:italic>in vitro<\/jats:italic>\n                    MCAD\u2010knockout (KO) HepG2 cells and a kinetic model of mFAO showed decreased CoASH, elevated medium\u2010chain acyl\u2010CoA, and decreased long\u2010chain acyl\u2010CoA levels. MCAD\u2010KO mice exposed to fasting and cold as energetic stressors had a significantly increased total CoA pool and increased expression of CoA biosynthetic enzymes in the liver, indicative of an upregulated CoA biosynthesis. Expression of carnitine acyltransferases and acyl\u2010CoA thioesterases, enzymes that liberate CoASH from acyl\u2010CoAs, was also upregulated, suggesting an adaptive response of CoA metabolism to decreased CoASH. Finally, computational model simulations showed that a combination of elevated total CoA and thioesterase activity led to normalisation of both CoASH and medium\u2010chain acyl\u2010CoA levels. Together, the results provide the first evidence for the CoA sequestration hypothesis in MCADD. The observed adaptation of CoA metabolism under energetic stress may act as a compensatory response that counteracts\u00a0CoASH depletion and accumulation of toxic medium\u2010chain acyl\u2010CoAs.\n                  <\/jats:p>","DOI":"10.1111\/febs.70442","type":"journal-article","created":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T08:06:34Z","timestamp":1770451594000},"page":"3565-3587","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Energetic stress in combination with impaired fatty acid oxidation induces sequestration of\n                    <scp>CoA<\/scp>\n                    and adaptation of\n                    <scp>CoA<\/scp>\n                    metabolism"],"prefix":"10.1111","volume":"293","author":[{"given":"Ligia Akemi","family":"Kiyuna","sequence":"first","affiliation":[{"name":"Laboratory of Paediatrics, University of Groningen University Medical Centre Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2199-5070","authenticated-orcid":false,"given":"Christoff","family":"Odendaal","sequence":"additional","affiliation":[{"name":"Laboratory of Paediatrics, University of Groningen University Medical Centre Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhulika","family":"Singh","sequence":"additional","affiliation":[{"name":"M Leiden Academic Centre for Drug Research, Leiden Academic Centre for Drug Research, Leiden University  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert","family":"Gerding","sequence":"additional","affiliation":[{"name":"Laboratory of Paediatrics, University of Groningen University Medical Centre Groningen  The Netherlands"},{"name":"Department of Laboratory Medicine University of Groningen, University Medical Centre Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam","family":"Langelaar\u2010Makkinje","sequence":"additional","affiliation":[{"name":"Laboratory of Paediatrics, University of Groningen University Medical Centre Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marianne","family":"van der Zwaag","sequence":"additional","affiliation":[{"name":"Biomedical Sciences\/ERIBA, University Medical Centre Groningen University of Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asmara","family":"Drachman","sequence":"additional","affiliation":[{"name":"M Leiden Academic Centre for Drug Research, Leiden Academic Centre for Drug Research, Leiden University  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladim\u00edra","family":"Cetkovsk\u00e1","sequence":"additional","affiliation":[{"name":"M Leiden Academic Centre for Drug Research, Leiden Academic Centre for Drug Research, Leiden University  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaby Liem Foeng","family":"Kioen","sequence":"additional","affiliation":[{"name":"M Leiden Academic Centre for Drug Research, Leiden Academic Centre for Drug Research, Leiden University  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne\u2010Claire M. 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