- high-resolution terminology - matching measurements at high-resolution
SUIT-002 O2 ce-pce D007
Description
Abbreviation: FNSGp(Oct,PGM)
Reference: A: SUIT 2 is the RP2 - SUIT-002
SUIT number: D007_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
O2k-Application: O2
- MitoPedia: SUIT - SUIT reference protocol RP2
- SUIT-category: FNSGp(Oct,PGM)
- SUIT protocol pattern: diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot
Harmonized SUIT protocols
RP1 and RP2
SUIT RP1
SUIT RP2
- SUIT RP1: 1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
- SUIT RP2: 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
Harmonization between RP1 and RP2
RP1 and RP2 in cells
RP2 can be assessed in cells after the selective plasma membrane permeabilization. Digitonin is a mild detergent that permeabilizes plasma membranes. The optimum effective digitonin concentration for complete plasma membrane permeabilization of cultured cells can be determined directly in a respirometric protocol (see Digitonin). The use of intact cells allows the possibility to evaluate respiration in two different modules: intact cells and permeabilized cells. ROUTINE respiration (ce1) is obtained before digitonin is employed, whereas after the use of digitonin we can address differentet Electron transfer-pathway statea.
- SUIT RP1 in cells: ce1;1Dig;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10Tm;11Azd
- SUIT RP2 in cells: ce1;1Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azdd
Harmonization between RP1 and RP2 in cells
RP1 and RP2 in PBMCs and PTLs
In PBMCs and PLTs the reference protocol RP1 is performed in the absense of Oct: SUIT 1 for PBMCs and PTLs
- SUIT RP1 in PBMCs and PTLsSUIT RP1 for PBMCs and PLTs:1PM;2D;2c;3U;4G;5S;6Rot;7Gp;8Ama;9AsTm;10Azd
- SUIT RP2 PBMCs and PLTs: 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
Harmonization between RP1 and RP2 in PBMC and PTL cells
References
Steps and respiratory states
Step
|
State
|
Pathway
|
Q-junction
|
Comment - Events (E) and Marks (M)
|
1D
|
REN
|
|
|
1D
- ADP is added to stimulate the consumption of endogenous fuel-substrates.
|
2M.1
|
|
|
|
1D;2M.1
|
3Oct
|
OctMP
|
F
|
FAO
|
1D;2M.1;3Oct
- Respiratory stimulation of the FAO-pathway, F, by fatty acid, FA, in the presence of malate, M. Malate is a type N substrate (N), required for the F-pathway. The FA concentration has to be optimized to saturate the F-pathway, without inhibiting or uncoupling respiration.
|
3c
|
OctMcP
|
F
|
FAO
|
1D;2M.1;3Oct;3c
- Respiratory stimulation of the FAO-pathway, F, by fatty acid, FA, in the presence of malate, M. Malate is a type N substrate (N), required for the F-pathway. The FA concentration has to be optimized to saturate the F-pathway, without inhibiting or uncoupling respiration.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
- Addition of cytochrome c yields a test for integrity of the mtOM (cytochrome c control efficiency). Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (OXPHOS capacity P with saturating [ADP]).
|
4M2
|
OctMP
|
F(N)
|
FAO
|
1D;2M.1;3Oct;3c;4M2
- Respiratory stimulation of the FAO-pathway, F, by fatty acid FA in the presence of malate M. Malate is a type N substrate (N), required for the F-pathway. In the presence of anaplerotic pathways (e.g., mitochondrial malic enzyme, mtME) the F-pathway capacity is overestimated, if there is an added contribution of NADH-linked respiration, F(N) (see SUIT-002). The FA concentration has to be optimized to saturate the FAO-pathway, without inhibiting or uncoupling respiration.
- High concentration of malate, typically 2 mM, saturates the N-pathway.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
5P
|
OctPMP
|
FN
|
FAO&CI
|
1D;2M.1;3Oct;4M2;5P
- Respiratory stimulation by simultaneous action of the F-pathway and N-pathway with convergent electron flow in the FN-pathway for evaluation of an additive or inhibitory effect of F.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
6G
|
OctPGMP
|
FN
|
FAO&CI
|
1D;2M.1;3Oct;4M2;5P;6G
- Respiratory stimulation by simultaneous action of the F-pathway and N-pathway with convergent electron flow in the FN-pathway for evaluation of an additive or inhibitory effect of F.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
7S
|
OctPGMSP
|
FNS
|
FAO&CI&II
|
1D;2M.1;3Oct;4M2;5P;6G;7S
- Respiratory stimulation by simultaneous action of the F-pathway, N-pathway, and S-pathway, with convergent electron flow in the FNS-pathway for reconstitution of TCA cycle function and additive or inhibitory effect of F.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
8Gp
|
OctPGMSGpP
|
FNSGp
|
FAO&CI&II&GpDH
|
1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp
|
9U
|
OctPGMSGpE
|
FNSGp
|
FAO&CI&II&GpDH
|
1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U
|
10Rot
|
SGpE
|
SGp
|
CII&GpDH
|
1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot
- Respiratory stimulation by action of succinate and glycerophosphate, Gp, with convergent electron flow in the SGp-pathway (CII&GpDH-linked pathway to the Q-junction).
- Noncoupled electron transfer state, ET state, with ET capacity E.
|
11Ama
|
ROX
|
|
|
1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama
- Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Β» Coupling control state
- Β» ET capacity
- Β» OXPHOS capacity
- Β» LEAK respiration
- 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
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Glossary
- Β» List of SUIT states
- Β» SUIT concept
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry
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