Step
|
Respiratory state
|
Pathway control
|
Pathway to Q
|
Comment
|
1OctM
|
OctM(L)
|
F
|
CETF
|
|
2D
|
OctM(P)
|
F
|
CETF
|
OXPHOS-coupling efficiency in state F.
|
3G
|
GMOct(P)
|
FN
|
FAO&CI
|
F/N ratio or (N-F)/N flux control factor of OXPHOS capacity.
|
4S
|
GMSOct(P)
|
FNS
|
FAO&CI&II
|
Additive effect of the NS pathway combination, compared to the sum of N+S pathway fluxes measured separately, in the OXPHOS state and the presence of F.
|
5Rot
|
S(P)
|
S
|
CII
|
The P/E ratio is not obtained in the FNS state, with preference given to the next steps for OXPHOS-coupling efficiency. A succinate concentration of >10 mM may be required for saturating SP capacity.
|
6Omy
|
S(L)
|
S
|
CII
|
OXPHOS-coupling efficiency in state S, comparable to state F.
|
7U
|
S(E)
|
S
|
CII
|
The complete ET capacity in state S may not be obtained after oligomycin. If apparent P/E>1.0, this is corrected to 1.0.
|
7c
|
S(E)
|
S
|
CII
|
If cytochrome c control factors = 0.0, the cytochrome c test conducted at this very late ET-pathway state provides an additional indication of stability of the mt-preparation during the experimental assay. Application of the cytochrome c test earlier in the protocol (SUIT-011: 1GM;2D;2c;3S; ..) is preferable, if cytochrome c control factors > 0.0 may be observed occasionally or regulary. If such cytochrome c effects are not exclusion criteria, then OXPHOS states with all substrate combinations can be compared in the presence of cytochrome c.
|
8Ama
|
|
ROX
|
ROX
|
ROX may be lower in substrate states earlier in the SUIT protocol. Therefore, this ROX measurement is frequently taken as a methodological control rather than as the basis of ROX correction of mitochondrial respiration (mt).
|