SUIT-004 O2 pfi D010: Difference between revisions
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{{MitoPedia | {{MitoPedia | ||
|abbr=NS(PM) | |abbr=NS(PM) | ||
|description=[[File:1PM;2D;2c;3U;4S;5Rot.png|300px]] | |description=[[File:1PM;2D;2c;3U;4S;5Rot.png|300px]][[File:SUIT_004.png|500px]] | ||
|SUIT number=D010_1PM;2D;3U;4S;5Rot;6Ama | |SUIT number=D010_1PM;2D;3U;4S;5Rot;6Ama | ||
|info='''A''': '''[[SUIT-004]]''' short protocol linked to [[SUIT-001 O2 pfi D002]] - '''SUIT RP1''' (human skeletal muscle) | |info='''A''': '''[[SUIT-004]]''' short protocol linked to [[SUIT-001 O2 pfi D002]] - '''SUIT RP1''' (human skeletal muscle) |
Revision as of 18:07, 24 January 2019
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;6Ama
O2k-Application: O2
- SUIT-category: NS(PM)
- SUIT protocol pattern: orthogonal 1PM;2D;2c;3U;4S;5Rot;6Ama
- Linked to SUIT-001 O2 pfi D002 - SUIT RP1, specifically for human skeletal muscle mitochondria.
- Harmonized with SUIT-005 O2 pfi D011.
Communicated by Iglesias-Gonzalez J and Gnaiger E (last update 2019-01-21)
References
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
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- A succinate concentration of >10 mM may be required for saturating SE capacity.
- + Linear coupling control (L-P-E) with N substrates (PM).
- + Pathway control in ET state (N, NS and S pathways).
- + 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 exper
Compare SUIT protocols
- The SUIT-004 protocol is linked to SUIT-001 O2 pfi D002 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,
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MitoPedia methods:
Respirometry