SUIT number: D011_1OctM;2D;3P;4S;5U;6Rot;7Ama;8AsTm;9Azd
SUIT-005 O2 pfi D011 protocols provide information on the F-pathway, the combined FN pathway, and the convergence FNS pathways in the OXPHOS state. FNS comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum OXPHOS- and ET-capacity. SUIT-005 can be extended with the CIV assay module. This protocol is linked to SUIT-002 O2 pfi D006 - SUIT RP2, specifically for human skeletal muscle mitochondria. SUIT-005 O2 pfi D011 is harmonized with SUIT-004 O2 pfi D010. The F-pathway has to be tested previously.
Communicated by Doerrier C, Iglesias-Gonzalez J and Gnaiger E (last update 2019-01-21)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)|
|Step||Respiratory state||Pathway control||ET-Complex entry into Q-junction||Comment|
- » Selected keywords in SUIT protocols
- Pathway control
Strengths and limitations
- + The protocol provides information on FAO capacity in the absence of other, potentially interfering pathways, both in the LEAK state and in OXPHOS.
- + FNS OXPHOS-capacity comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
- + FNS ET-capacity is a good estimate of overall ET-capacity in many cell types.
- + The presence of PM and S establishes a fully operative TCA cycle activity (PMS) if the 2-oxoglutarate carrier does not outcompete the sources of 2-oxoglutarate
- + Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.
- + Reasonable duration of the experiment (1.5 to 2 h); 2-3 h when the CIV module is added.
- - F- OXPHOS-capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemuieux et al 2011).
- - SRotE may be underestimated if S is not saturating.
Compare SUIT protocols
- SUIT-002: A comparable but more comprehensive protocol comprising additional substrate states.
- SUIT-002 O2 pfi D006: A comparable but more comprehensive pfi-specific protocol comprising additional substrate states.
- imt, ROX: 1OctM;2D;2c;3P;4S;5U;6Rot;7Ama
- pfi, high O2, no ROX: FNS(PM)01_pfiO2,1OctM,2D(c),3P,4S,5U,6Rot
MitoPedia methods: Respirometry