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Template:SUIT-001

From Bioblast

SUIT-001

Step Respiratory state Pathway control ET-Complex entry into Q-junction Comment
1PM PML N CI PML or PM_L: Pyruvate & Malate, N-LEAK respiration, NL

NADH-linked substrates (type N; CI-linked pathway to Q). Non-phosphorylating resting state (LEAK state); Ln in the absence of ADP, ATP, AMP (no adenylates).

2D PMP N CI PMP or PM_P: N-OXPHOS capacity, NP

OXPHOS capacity, P (with saturating [ADP]), with NADH-linked substrates.

2c PMcP N CI PMcP or PMc_P: Cytochrome c test for quality control

Addition of cytochrome c yields a test for integrity of the mtOM. Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in state NP due to loss of cytochrome c (P, OXPHOS capacity with type N substrates). Cytochrome c is added immediately after the earliest ADP-activation step.

3U PME N CI PME or PM_E: N-ET capacity, NE

Uncoupler titration (avoiding inhibition by high uncoupler concentrations) to obtain electron transfer (ET) capacity (noncoupled ET-state), as a test for limitation of OXPHOS-capacity by the phosphorylation system (ANT, ATP synthase, phosphate transporter) relative to ET-capacity, E.

4G PGM(E) N CI
5S PGMSE NS CI&II GMSP or GMS_P: NS-OXPHOS capacity, NSP

Respiratory stimulation by further addition of succinate, S, to type N substrates, with convergent electron flow in the NS-pathway (CI&II-linked pathway to the Q-junction) for reconstitution of TCA cycle function, in the coupled state as an estimate of OXPHOS-capacity, P.

6Oct OctPGMS(E) FNS FAO&CI&II
7Rot SE S CII SE or S_E: S-ET capacity

S-pathway ET-capacity after blocking CI with rotenone, inhibiting the N- and F-pathways.

8Gp SGp(E) SGp CII&GpDH
9Ama ROX ROX: residual oxygen consumption

Rox is due to oxidative side reactions, estimated after addition of Antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).