Difference between revisions of "SUIT-001 O2 mt D001"
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::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp | ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp | ||
__TOC__ | __TOC__ | ||
{{Template:SUIT-001}} | {{Template:SUIT-001}} | ||
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:::- The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring S<sub>''E''</sub>. | :::- The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring S<sub>''E''</sub>. | ||
:::- Lengthy duration of the experiment. | :::- Lengthy duration of the experiment. | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
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::::* [[SUIT-006]]: Very short initial coupling control module. | ::::* [[SUIT-006]]: Very short initial coupling control module. | ||
== References == | |||
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{{MitoPedia concepts | {{MitoPedia concepts | ||
|mitopedia concept=SUIT protocol, SUIT A, Find | |mitopedia concept=SUIT protocol, SUIT A, Find |
Revision as of 12:23, 20 February 2019
Description
Abbreviation: FNSGp(Oct,PGM)
Reference: A Reference protocol RP1; mt: isolated mitochondria and tissue homogenate - SUIT-001
SUIT number: D001_1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
O2k-Application: O2
- MitoPedia: SUIT - SUIT reference protocol RP1 for isolated mitochondria and tissue homogenate
- SUIT-category: FNSGp(Oct,PGM)
- SUIT protocol pattern: orthogonal 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp
Steps and respiratory states
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
|
3U | PME | N | CI | 1PM;2D;3U
|
4G | PGME | N | CI | 1PM;2D;3U;4G
|
5S | PGMSE | NS | CI&II | 1PM;2D;3U;4G;5S
|
6Oct | OctPGMSE | FNS | FAO&CI&II | 1PM;2D;3U;4G;5S;6Oct
|
7Rot | SE | S | CII | 1PM;2D;3U;4G;5S;6Oct;7Rot
|
8Gp | SGpE | SGp | CII&GpDH | 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp
|
9Ama | ROX | 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp;9Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
- 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-001 in combination with SUIT-002 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.
- + Linear coupling control (L-P-E) with N substrates (PM).
- + Pathway control in ET state (N, NS, FNS, S and SGp pathways).
- + Harmonization with SUIT-002 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated measurements.
- + In mtprep, SUIT-001 and SUIT-002 harmonize in: SGpE (8Gp in SUIT-001 and 10Rot in SUIT-002) and Complex IV.
- + Harmonization with many SUIT protocols (up to step 5S).
- + Oct is added in ET state to avoid uncoupler effect with fatty acids.
- + If NS- and FNS-pathways are similar (NSE ~ FNSE), the additivity index of N- and S-pathways in ET state can be evaluated.
- + This protocol can be extended with the Complex IV module.
- - The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring SE.
- - Lengthy duration of the experiment.
Compare SUIT protocols
- SUIT-002 O2 mt D005 (RP2): 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, 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 |
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry
Labels:
SUIT-001