From Bioblast
Description
Abbreviation: NS(PM)
Reference: A: Short protocol linked to SUIT-001 O2 pfi D002 - SUIT RP1 (human skeletal muscle)-SUIT-004
SUIT number: D010_1PM;2D;2c;3U;4S;5Rot;6Ama;7AsTm;8Azd
O2k-Application: O2
- MitoPedia: SUIT
- 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.
The SUIT-004 O2 pfi D010 protocol provides information of the linear coupling control (L- P- E) with NADH linked-substrates (PM). Moreover, the pathway control in ET state (N, NS and S) can be evaluated by using this SUIT protocol. The SUIT-004 O2 pfi D010 can be extended with the CIV assay module.
Communicated by 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) |
---|---|---|---|---|
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
- A succinate concentration of >10 mM may be required for saturating SE capacity.
- CIV assessment is skipped if a prolonged protocol is not practical due to time limitation.
- RP1-6Ama may be skipped since correction of the initial states by ROX measured in RP1-6 Ama may represent an over-correction, and inhibition takes a very long time in several cases (Pesta 2012 Methods Mol Biol).
- + Linear coupling control (L- P- E) with NADPH-linked substrate substrates (PM).
- + Pathway control in ET state (N, NS, S).
- + This protocol can be extended with the Complex IV module.
- + Short experimental time.
- - The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-004 in favor of reducing the time consumed during the experiment.
Compare SUIT protocols
- SUIT-001 O2 pfi D002: A comparable but more comprehensive protocol comprising additional substrate states.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry