From Bioblast
Description
Abbreviation: FNSGp(Oct,PGM)
Reference: A Reference protocol RP2; mt: isolated mitochondria and tissue homogenate - SUIT-002
SUIT number: D005_1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
O2k-Application: O2
- MitoPedia: SUIT - SUIT reference protocol RP2 for isolated mitochondria and tissue homogenate
- SUIT-category: FNSGp(Oct,PGM)
- SUIT protocol pattern: diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot-
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet21.06 SUIT RP | 2018-06-25 | SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. | ||
Doerrier 2018 Methods Mol Biol | 2018 | Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, MΓ©szΓ‘ros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. https://doi.org/10.1007/978-1-4939-7831-1_3 | Human Mouse Rat Saccharomyces cerevisiae | Heart Skeletal muscle Endothelial;epithelial;mesothelial cell Blood cells HEK Platelet |
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1D | REN | 1D
| ||
2M.1 | 1D;2M.1 | |||
3Oct | OctMP | F | FAO | 1D;2M.1;3Oct
|
3c | OctMcP | F | FAO | 1D;2M.1;3Oct;3c
|
4M2 | OctMP | F(N) | FAO | 1D;2M.1;3Oct;3c;4M2
|
5P | OctPMP | FN | FAO&CI | 1D;2M.1;3Oct;4M2;5P
|
6G | OctPGMP | FN | FAO&CI | 1D;2M.1;3Oct;4M2;5P;6G
|
7S | OctPGMSP | FNS | FAO&CI&II | 1D;2M.1;3Oct;4M2;5P;6G;7S
|
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
|
11Ama | ROX | 1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama
|
- 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
- SUIT-002 in combination with SUIT-001 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.
- + SUIT-002 allows the depletion of endogenous substrates with ADP (1D).
- + The protocol provides information on F-pathway in OXPHOS state. The low concentration of malate used in this protocol, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
- + F-pathway (3Oct-2M.1) compared to FN-pathway (5P) in OXPHOS state.
- + Pathway control in OXPHOS (F, F(N), FN, FNS, FNSGp pathways) and in ET state (FNSGp and SGp).
- + Harmonization with SUIT-001 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statically used as repeat measurements.
- + Harmonization with many SUIT protocols (up to step 7S).
- + In SUIT-002, the full set of pathways converging into Q (FNSGp) is obtained in OXPHOS and ET states. Therefore, P/E (8Gp/9U) at high ET capacity can be calculated.
- + This protocol can be extended with the Complex IV module.
- - S-pathway in ET state is not obtained (it is obtained in SUIT-001).
- - Lengthy duration of the experiment.
Compare SUIT protocols
- SUIT-001 O2 mt D001 (RP1): Harmonized SUIT protocol for isolated mitochondria and tissue homogenate.
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry
Labels:
SUIT-002