1. Oxidative phosphorylation analysis: assessing the integrated functional activity of human skeletal muscle mitochondria--case studies
- Author
-
Neil R. Friedman, Douglas S. Kerr, Charles L. Hoppel, Marie E. Varnes, Bruce H. Cohen, and Michelle Puchowicz
- Subjects
Fatty acid metabolism ,Adenine nucleotide translocator ,Cell Biology ,Oxidative phosphorylation ,Mitochondrion ,Biology ,Pyruvate dehydrogenase complex ,chemistry.chemical_compound ,chemistry ,Biochemistry ,medicine ,biology.protein ,Molecular Medicine ,Carnitine ,Acetylcarnitine ,Molecular Biology ,Palmitoylcarnitine ,medicine.drug - Abstract
Oxidative phosphorylation analysis, performed on freshly-isolated mitochondria, assesses the integrated function of the electron transport chain (ETC) coupled to ATP synthesis, membrane transport, dehydrogenase activities, and the structural integrity of the mitochondria. In this review, a case study approach is employed to highlight detection of defects in the adenine nucleotide translocator, the pyruvate dehydrogenase complex, fumarase, coenzyme Q function, fatty acid metabolism, and mitochondrial membrane integrity. Our approach uses the substrates glutamate, pyruvate, 2-ketoglutarate (coupled with malonate), malate, and fatty acid substrates (palmitoylcarnitine, octanoylcarnitine, palmitoyl-CoA (with carnitine), octanoyl-CoA (with carnitine), octanoate and acetylcarnitine) in addition to succinate, durohydroquinone and TMPD/ascorbate to uncover metabolic defects that would not be apparent from ETC assays performed on detergent-solubilized mitochondria.
- Published
- 2003