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. 2020 Oct 19;10:17599. doi: 10.1038/s41598-020-74718-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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The mitochondrial SLC family of proteins correlate with Pyr/M supported intrinsic phosphorylation efficiency. (A) Respiration in isolated mitochondria energized with Pyr/M. Respiration was stimulated with ΔG_ATP of − 54.16 kJ/mol (CK), leak respiration in the absence of adenylates (S4), 0.01 mM ADP (ADP_10µM), 0.5 mM ADP (ADP_500µM), and FCCP in the presence of 0.5 mM ADP (FCCP_HK), no adenylates (FCCP_Alone), or ΔG_ATP (FCCP_CK). (B) Relationship between mitochondrial ATP synthesis and oxygen consumption, a direct measurement of intrinsic phosphorylation efficiency (i.e., P/O ratio). Substrate condition = Pyr/M. Dashed line depicts the theoretical maximum for P/O based on the known mammalian stoichiometry of ATP synthesis. (C) Respiration in isolated mitochondria energized with succinate/rotenone. Respiration was stimulated with ΔG_ATP of − 54.16 kJ/mol (CK), leak respiration in the absence of adenylates (S4), 0.01 mM ADP (ADP_10µM), 0.5 mM ADP (ADP_500µM), and FCCP in the presence of 0.5 mM ADP (FCCP_HK), no adenylates (FCCP_Alone), or ΔG_ATP (FCCP_CK). (D) Relationship between mitochondrial ATP synthesis and oxygen consumption, a direct measurement of intrinsic phosphorylation efficiency (i.e., P/O ratio). Substrate condition = succinate/rotenone. Dashed line depicts the theoretical maximum for P/O based on the known mammalian stoichiometry of ATP synthesis. (E) Correlation of MitoCarta 2.0 normalized protein abundance of Ucp3, Lyrm7, and Scl25a46 with Pyr/M supported P/O. (F) MitoCarta 2.0 normalized protein expression of identified/quantified mitochondrial SLC proteins. Stars indicate SLC proteins found to correlate with Pyr/M mediated P/O. Figures generated using GraphPad Prism 8 software (Version 8.4.2). Data information: Data are Mean ± SEM, N = 5/group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.