Difference between revisions of "SUIT-002"
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNSGp(Oct,PGM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNSGp(Oct,PGM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1D;2M;3Oct;4M;5P;6G;7S;8Gp;9U;10Rot- | ::: '''[[SUIT protocol pattern]]:''' diametral 1D;2M;3Oct;4M;5P;6G;7S;8Gp;9U;10Rot- | ||
The SUIT-002 protocol in combination with [[SUIT-001]] provides a common reference for comparison of respiratory control of [[Mitochondrial preparations| mitochondrial preparations]] in a wide variety of species, tissues and cell types. SUIT-002 is specially designed to give information on F-pathway in OXPHOS state avoiding FAO overestimation in the presence of anaplarotic pathways. Moreover, the pathway control in OXPHOS state (F, F(N), FN, FNS, FNSGp pathways) and in ET state (FNSGp and SGp) can be evaluated by using this SUIT protocol. SUIT-002 can be extended with the CIV assay module. | |||
__TOC__ | __TOC__ | ||
Communicated by [[Doerrier C]] and [[Gnaiger E]] (last update 2019-01-28) | Communicated by [[Doerrier C]] and [[Gnaiger E]] (last update 2019-01-28) |
Revision as of 14:15, 30 January 2019
Description
Abbreviation: FNSGp(Oct,PGM)
Reference: A: RP2 Doerrier 2018 Methods Mol Biol »Versions
- SUIT-category: FNSGp(Oct,PGM)
- SUIT protocol pattern: diametral 1D;2M;3Oct;4M;5P;6G;7S;8Gp;9U;10Rot-
The SUIT-002 protocol in combination with SUIT-001 provides a common reference for comparison of respiratory control of mitochondrial preparations in a wide variety of species, tissues and cell types. SUIT-002 is specially designed to give information on F-pathway in OXPHOS state avoiding FAO overestimation in the presence of anaplarotic pathways. Moreover, the pathway control in OXPHOS state (F, F(N), FN, FNS, FNSGp pathways) and in ET state (FNSGp and SGp) can be evaluated by using this SUIT protocol. SUIT-002 can be extended with the CIV assay module.
Communicated by Doerrier C and Gnaiger E (last update 2019-01-28)
DLP applications
- SUIT-002 O2 mt D005 for isolated mitochondria and tissue homogenate
- SUIT-002 O2 pfi D006 for permeabilized fibers
- SUIT-002 O2 pce D007 for permeabilized cells
- SUIT-002 O2 pce D007a for permeabilized PBMCs and PLTs (absense of Oct)
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Doerrier 2018 Methods Mol Biol | 2018 | Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros 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
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » 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).
Compare SUIT protocols
MitoPedia concepts:
MiP concept,
SUIT protocol,
Recommended
MitoPedia methods:
Respirometry