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| [[Electron-transfer-pathway state]] will depend on the substrate added to fuel the mitochondrial respiration.
| | ::::» [[Entity]] ''X'' |
| To obtain the '''NADH electron-transfer-pathway state''' (N), [[NADH]]-linked substrates (CI-linked) are added, feeding electrons into the [[N-junction]] catalyzed by various mt-dehydrogenases. The most commonly applied N-junction substrate combinations are: [[PM]], [[GM]], [[PGM]]. The '''Succinate pathway control state''' (S), succinate-linked respiration or S-pathway is achieved with '''[[succinate]]''' as the single substrate. It is recommended to investigate the S-linked respiration in the presence of [[rotenone]] (Rot; or amytal, piericidine), which prevents accumulation of [[oxaloacetate]]. Oxaloacetate is a potent inhibitor of [[Complex II]] (CII; see [[succinate dehydrogenase]], SDH). After inhibition of [[Complex I]] by rotenone, the NADH-linked dehydrogenases become inhibited by the redox shift from NAD<sup>+</sup> to NADH. SDH is activated by succinate and ATP, which explains in part the time-dependent increase of respiration in isolated mitochondria after addition of rotenone (first), succinate and ADP.
| | ::::» [[Elementary unit]] x |
| :The following substrates can also be applied in this protocol: | |
| * [[Glycerophosphate]] (Gp) which feeds the Q-junction
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| * Fatty acids to obtain the [[Fatty acid oxidation pathway control state]]
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