SUIT-001 O2 ce-pce D003: Difference between revisions
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::: [[MitoPedia: SUIT]] - '''[[SUIT reference protocol]] RP1 for permeabilized cells''' | ::: [[MitoPedia: SUIT]] - '''[[SUIT reference protocol]] RP1 for permeabilized cells''' | ||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNSGp(Oct,PGM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNSGp(Oct,PGM) | ||
::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp | ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp- | ||
__TOC__ | __TOC__ | ||
Revision as of 15:26, 20 February 2019
Description
Abbreviation: FNSGp(Oct,PGM)
Reference: A Reference protocol RP1; pce: permeabilized cells - SUIT-001
SUIT number: D003_ce1;1Dig;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
O2k-Application: O2
- MitoPedia: SUIT - SUIT reference protocol RP1 for permeabilized cells
- SUIT-category: FNSGp(Oct,PGM)
- SUIT protocol pattern: orthogonal 1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp-
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
1Dig | REN | ce1;1Dig
|
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
|
4G | PGME | N | CI | 1PM;2D;3U;4G
|
5S | PGMSE | NS | CI&II | 1PM;2D;3U;4G;5S
|
6Oct | OctPGMSE | FNS | FAO&CI&II | 1PM;2D;3U;4G;5S;6Oct
|
7Rot | SE | S | CII | 1PM;2D;3U;4G;5S;6Oct;7Rot
|
8Gp | SGpE | SGp | CII&GpDH | 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp
|
9Ama | ROX |
|
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
- SUIT-001 in combination with SUIT-002 provides a common reference for comparison of respiratory control in a large variety of species, tissues and cell types. Both SUIT protocols provide a mitochondrial mapping which allows:
- 1. to obtain reference values.
- 2. to evaluate mitochondrial physiological diversity, generating a mt-database on comparative mitochondrial physiology.
- 3. to screen specific defects.
- SUIT-001 and SUIT-002 are used in the MitoFit Proficiency test for inter-individual and inter-laboratory reproducibility quality control.
- 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, FNS, S and SGp pathways).
- + Harmonization with SUIT-002 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated measurements.
- + In cells, SUIT-001 and SUIT-002 harmonize in: ROUTINE (ce1), SGpE (8Gp in SUIT-001 and 10Rot in SUIT-002) and Complex IV.
- + Harmonization with many SUIT protocols (up to step 5S).
- + Oct is added in ET state to avoid uncoupler effect with fatty acids.
- + If NS- and FNS-pathways are similar (NSE ~ FNSE), the additivity index of N- and S-pathways in ET state can be evaluated.
- + This protocol can be extended with the Complex IV module.
- - The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring SE.
- - Lengthy duration of the experiment.
Compare SUIT protocols
- SUIT-001 O2 pce D003 (general SUIT-001 for permeabilized cells) differs from SUIT-001 O2 pce D004 (SUIT-001 for permeabilized PBMCs and PLTs) in the presence and absence of octanoylcarnitine, respectively.
- SUIT-002 O2 pce D007 (RP2): Harmonized SUIT protocol for permeabilized cells.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry
Labels:
SUIT-001