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Respirometry with living cells provides limited information compared to precision OXPHOS analysis with mitochondrial preparations, including isolated mitochondria, tissue homogenates, permeabilized tissues, and permeabilized cells. We studied mouse mitochondria from brain, a glucose\u2010dependent tissue, and from heart, which relies highly on fatty acid oxidation (FAO). HEK 293T cells were analysed as a widely used experimental model. Human peripheral blood mononuclear cells (PBMCs) and platelets were obtained from non\u2010invasive liquid biopsies, considering their potential as mitochondrial biomarkers. Twenty respiratory states were interrogated applying two substrate\u2013uncoupler\u2013inhibitor titration (SUIT) reference protocols in parallel. Convergent electron transfer (ET) into the coenzyme Q junction increased OXPHOS and ET capacities compared to separately stimulated\u00a0pathways. In mouse heart and human PBMCs, OXPHOS capacities were identical to ET capacities in every pathway state. While this equivalence applied to the NADH\u2010linked pathway in platelets, ET capacity exceeded OXPHOS capacity supported by NADH\u2010linked substrates plus succinate. Surprisingly, mouse brain exhibited the highest excess ET capacity in the NADH\u2010linked pathway. In contrast, ET capacity of different batches of HEK 293T cells varied at constant OXPHOS capacity. Precision OXPHOS analysis enables attribution of respiratory performance to nutrient\u2010specific pathways. In studies ranging from exercise physiology to mitochondrial diseases, metabolic adjustments must be distinguished from functional defects. Bioenergetic profiles obtained by precision OXPHOS analysis gain perspective in the context of comparative mitochondrial physiology.<\/jats:p>","DOI":"10.1113\/ep092792","type":"journal-article","created":{"date-parts":[[2025,8,1]],"date-time":"2025-08-01T12:10:32Z","timestamp":1754050232000},"page":"179-211","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Bioenergetic profiles and respiratory control in mitochondrial physiology: Precision analysis of oxidative phosphorylation"],"prefix":"10.1113","volume":"111","author":[{"given":"Alba","family":"Tim\u00f3n\u2010G\u00f3mez","sequence":"first","affiliation":[{"name":"Oroboros Instruments  Innsbruck Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina","family":"Doerrier","sequence":"additional","affiliation":[{"name":"Oroboros Instruments  Innsbruck Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzana","family":"Sumbalov\u00e1","sequence":"additional","affiliation":[{"name":"Oroboros Instruments  Innsbruck Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luiz F.","family":"Garcia\u2010Souza","sequence":"additional","affiliation":[{"name":"Oroboros Instruments  Innsbruck Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eleonora","family":"Baglivo","sequence":"additional","affiliation":[{"name":"Oroboros Instruments  Innsbruck Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luiza H. 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