SUIT-031 O2 ce-pce D079: Difference between revisions
From Bioblast
No edit summary Tag: Reverted |
No edit summary Tag: Manual revert ย |
||
(One intermediate revision by the same user not shown) | |||
Line 28: | Line 28: | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
:::* [[SUIT-001 O2 | :::* [[SUIT-001 O2 mt D001]] is an extended coupling-control proton in the N-pathway and contribution of G-, S-, Oct- and Gp-pathways in the ET state for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k-chamber. ย | ||
:::* [[SUIT-004]] is designed to provide a quick assessment of the linear coupling control (L- P- E) with NADH-linked substrates (PM) and the contribution of the S-pathway to the ET state (N, NS, S). ย | :::* [[SUIT-004]] is designed to provide a quick assessment of the linear coupling control (L- P- E) with NADH-linked substrates (PM) and the contribution of the S-pathway to the ET state (N, NS, S). ย | ||
:::* [[SUIT-006 O2 mt D047]] is a coupling-control protocol in the N-pathway for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k-chamber. | :::* [[SUIT-006 O2 mt D047]] is a coupling-control protocol in the N-pathway for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k-chamber. |
Latest revision as of 12:15, 2 December 2021
Description
Abbreviation: PM+S+Rot
Reference: A to analyse O2 flux in the NS- pathway control state in permeabilized cells- SUIT-031
SUIT number: D079_ce1;1Dig;1PM;2D;2c;3S;4Rot;5U;6Ama
O2k-Application: O2
This is a protocol to investigate oxygen flux N- and NS-pathway and S-pathway control state in the OXPHOS state and S-pathway control state in the ET state in permeabilized cells. SUIT-031 Q mt D072 and SUIT-031 Q ce-pce D074 are designed to measure simultaneously O2 flux and the Q-redox state in isolated mitochondria and permeabilized cells.
Communicated by Komlodi T (last update 2021-12-02)
Representative traces
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
|
3S | PMSP | NS | CI&II | 1PM;2D;2c;3S
|
4Rot | SP | S | CII | 1PM;2D;2c;3S;4Rot
|
5U | SE | N | CII | 1PM;2D;2c;3S;4Rot;5U
|
6Ama | 1PM;2D;2c;3S;4Rot;5U;6Ama
|
- 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-031 O2 mt D075 in combination with SUIT-006 O2 mt D022 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.
- + This protocol can be extended with the Complex IV module.
- + Reasonable duration of the experiment.
- - Careful washing is required after the experiment to avoid carry-over of uncoupler and inhibitors.
Compare SUIT protocols
- SUIT-001 O2 mt D001 is an extended coupling-control proton in the N-pathway and contribution of G-, S-, Oct- and Gp-pathways in the ET state for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k-chamber.
- SUIT-004 is designed to provide a quick assessment of the linear coupling control (L- P- E) with NADH-linked substrates (PM) and the contribution of the S-pathway to the ET state (N, NS, S).
- SUIT-006 O2 mt D047 is a coupling-control protocol in the N-pathway for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k-chamber.
- SUIT-008 is designed to assess the additivity between the N- and S-pathway in the Q-junction, providing a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1Dig | Digitonin (Dig) |
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1PM | Pyruvate (P) and Malate (M) | |
2D | ADP (D) | |
2c | Cytochrome c (c) | |
3S | Succinate (S) | |
4Rot | Rotenone (Rot) | |
5U | Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U) | Can be substituted for other uncoupler. |
6Ama | Antimycin A (Ama) |
- Suggested stock concentrations are shown in the specific DL-Protocol.
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet24.12 NextGen-O2k: Q-Module | 2021-10-29 | NextGen-O2k: Q-Module manual |
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry