SUIT-002 O2 mt D005: Difference between revisions
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|SUIT number=D005_1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd Β | |SUIT number=D005_1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd Β | ||
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::: [[MitoPedia: SUIT]] - '''[[SUIT reference protocol]] RP2 for isolated mitochondria and tissue homogenate''' | ::: '''[[MitoPedia: SUIT]]''' - '''[[SUIT reference protocol]] RP2 for isolated mitochondria and tissue homogenate''' | ||
::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNSGp(Oct,PGM) | ::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNSGp(Oct,PGM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot- | ::: '''[[SUIT protocol pattern]]:''' diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot- | ||
The SUIT-002 O2 mt D005 protocol in combination with [[SUIT-001_O2_mt_D001]] 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 O2 mt D005 is specially designed to give information on [[Fatty_acid_oxidation_pathway_control_state|F-pathway]] in [[Oxidative phosphorylation|OXPHOS state]] avoiding FAO overestimation in the presence of [[Anaplerosis|anaplerotic]] pathways. Moreover, the pathway control in OXPHOS state ([[Fatty_acid_oxidation_pathway_control_state|F]], F(N), [[FN]], [[FNS]], [[FNSGp]] pathways) and in [[ET-capacity| ET state]] ([[FNSGp]] and [[SGp-pathway control state|SGp]]) can be evaluated by using this SUIT protocol. SUIT-002 O2 mt D005 can be extended with the CIV assay module. | The SUIT-002 O2 mt D005 protocol in combination with [[SUIT-001_O2_mt_D001]] provides a common reference for comparison of respiratory control of [[Mitochondrial preparations| mitochondrial preparations]] (isolated mitochondria and tissue homogenates) in a wide variety of species, tissues and cell types. SUIT-002 O2 mt D005 is specially designed to give information on [[Fatty_acid_oxidation_pathway_control_state|F-pathway]] in [[Oxidative phosphorylation|OXPHOS state]] avoiding FAO overestimation in the presence of [[Anaplerosis|anaplerotic]] pathways. Moreover, the pathway control in OXPHOS state ([[Fatty_acid_oxidation_pathway_control_state|F]], F(N), [[FN]], [[FNS]], [[FNSGp]] pathways) and in [[ET-capacity| ET state]] ([[FNSGp]] and [[SGp-pathway control state|SGp]]) can be evaluated by using this SUIT protocol. SUIT-002 O2 mt D005 can be extended with the CIV assay module. | ||
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Communicated by [[Doerrier C]] and [[Gnaiger E]] (last update 2019-03-06) | |||
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== References == | == References == | ||
{{#ask:[[Category:Publications]] [[Additional label::SUIT-002 O2 mt D005]] | {{#ask:[[Category:Publications]] [[Additional label::SUIT-002 O2 mt D005]] [[Additional label::SUIT-002]] | ||
| ?Was published in year=Year | | ?Was published in year=Year | ||
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|mitopedia method=Respirometry | |mitopedia method=Respirometry | ||
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Revision as of 13:50, 6 March 2019
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-
The SUIT-002 O2 mt D005 protocol in combination with SUIT-001_O2_mt_D001 provides a common reference for comparison of respiratory control of mitochondrial preparations (isolated mitochondria and tissue homogenates) in a wide variety of species, tissues and cell types. SUIT-002 O2 mt D005 is specially designed to give information on F-pathway in OXPHOS state avoiding FAO overestimation in the presence of anaplerotic 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 O2 mt D005 can be extended with the CIV assay module.
Communicated by Doerrier C and Gnaiger E (last update 2019-03-06)
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) can be 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) can be observed.
- + Harmonization with SUIT-001 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated 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.
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 |
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry