The SUIT-005 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.
Communicated by Iglesias-Gonzalez J, Krumschnabel G and Gnaiger E (last update 2019-01-26)
Specific SUIT protocols
SUIT-005 O2 pfi D11
- SUIT-005 O2 pfi D011 for permeabilized fibers
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
- + Reasonable duration of the experiment (1.5 to 2 h); 2-3 h when the CIV module is added.
- + The protocol provides information on F-OXPHOS capacity in the absence of a significant OXPHOS-capacity in the malate anaplerotic pathway control state.
- + FNS comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum OXPHOS- and ET-capacity.
- + PM & S reconstitutes 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.
- - F-OXPHOS capacity may be underestimated with Oct. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) & malate increased OXPHOS-capacity by 26% (Lemuieux et al 2011).
- - SRotE may be underestimated if S is not saturating.
Compare SUIT protocols
- SUIT-002 check for malate-linked anaplerotic activity
MitoPedia methods: Respirometry