From Bioblast
Description
Abbreviation: NS(PM)
Reference: A: SUIT-004 short protocol linked to SUIT-001 O2 pfi D002 - SUIT RP1 (human skeletal muscle)
SUIT number: D010_1PM;2D;3U;4S;5Rot
O2k-Application: O2
- SUIT-category: NS(PM)
- SUIT protocol pattern: orthogonal 1PM;2D;2c;3U;4S;5Rot
- Linked to SUIT-001 O2 pfi D002 - SUIT RP1, specifically for human skeletal muscle mitochondria.
- Harmonized with SUIT-005 O2 pfi D011.
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet21.06 SUIT RP | 2018-06-25 | SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. | ||
Doerrier 2018 Methods Mol Biol | 2018 | Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, MΓ©szΓ‘ros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. https://doi.org/10.1007/978-1-4939-7831-1_3 | Human Mouse Rat Saccharomyces cerevisiae | Heart Skeletal muscle Endothelial;epithelial;mesothelial cell Blood cells HEK Platelet |
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1PM | PML(n) | N | CI | 1PM
|
2D | PMP | N | CI | 1PM;2D
|
2c | PMcP | N | CI | 1PM;2D;2c
|
3U | PME | N | CI | 1PM;2D;3U
|
4S | PMSE | NS | CI&II | 1PM;2D;3U;4S
|
5Rot | SE | S | CII | 1PM;2D;3U;4S;5Rot
|
6Ama | ROX | 1PM;2D;3U;4S;5Rot;6Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
Compare SUIT protocols
Link to RP1
- The 1PM;2D;3U;4S;5Rot- protocol is linked to 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp- for healthy human skeletal muscle mitochondria on the basis of the following rationale.
- The first coupling control steps (1-4) are identical.
- RP1-5G is skipped, since PM and PGM yield nearly identical OXPHOS capacity. However, PM<PGM is a potential diagnostic for a defect in the PM-linked pathway upstream of CI.
- RP1-7Oct is skipped, since octanoylcarnitine exerts practically no stimulatory effect on OXPHOS capacity in state PMS or PGMS, but in conjunction with prolonged exposure may even yield a slight decline of respiration.
- RP1-9Gp is skipped, since glycerolphosphate exerts only a minor stimulatory effect on OXPHOS capacity in state SRot.
- Omission of the three steps described above shortens the SUIT protocol, thus reducing the risk of diminishing respiratory capacity over time, reducing the number of necessary reoxygenations, and providing more time for detailed evaluation of steady-states. The loss of diagnostic information may be minor relative to the benefits of simplification of the protocol, particularly in studies of exercise in healthy humans.
- RP1-10Ama may be skipped, since correction of the initial states by ROX measured in RP1-10 Ama may represent an over-correction, and inhibition takes a very long time in several cases (Pesta 2012 Methods Mol Biol).
- RP1-11Tm and RP1-12Azd are skipped, if a prolonged protocol is not practical due to limination of time.
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry