Difference between revisions of "SUIT-012"
From Bioblast
Line 1: | Line 1: | ||
{{MitoPedia | {{MitoPedia | ||
|abbr=N(PGM) | |abbr=N(PGM) | ||
|description=[[File:1PM;2D;3G;4U-.png|300px]] | |description=[[File:1PM;2D;3G;4U-.png|300px]] | ||
|info='''A''' [[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/6/64/SUIT-012.pdf|Bioblast pdf]] »[http://wiki.oroboros.at/index.php/File:SUIT-012.pdf Versions] | |info='''A''' [[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/6/64/SUIT-012.pdf|Bioblast pdf]] »[http://wiki.oroboros.at/index.php/File:SUIT-012.pdf Versions] | ||
}} | }} | ||
{{MitoPedia concepts | |||
|mitopedia concept=MiP concept, SUIT protocol, Recommended | |||
}} | |||
{{MitoPedia methods | |||
|mitopedia method=Respirometry | |||
}} | |||
{{MitoPedia O2k and high-resolution respirometry}} | |||
{{MitoPedia topics}} | |||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' N(PGM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' N(PGM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1PM;2D;2c;3G;4U- | ::: '''[[SUIT protocol pattern]]:''' diametral 1PM;2D;2c;3G;4U- | ||
Line 34: | Line 42: | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::+ This protocol allows to evaluate the function of the TCA cycle without the involvement of the [[complex II]] (S-pathway). | :::+ This protocol allows to evaluate the function of the TCA cycle without the involvement of the [[complex II]] (S-pathway). It is thus useful to understand the contribution and the activity of dehydrogenases providing NADH. | ||
:::+ Mitochondrial external membrane can be measured with the addition of [[cytochrome c]]. Application of the [[cytochrome c]] test early in the protocol ensures comparability of all states in case of any effect of c. | :::+ Mitochondrial external membrane integrity can be measured with the addition of [[cytochrome c]]. Application of the [[cytochrome c]] test early in the protocol ensures comparability of all states in case of any effect of c. | ||
:::+ Reasonable duration of the experiment. | :::+ Reasonable duration of the experiment. | ||
:::+ Complex IV activity can be measured. | :::+ Complex IV activity can be measured. | ||
Line 43: | Line 51: | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
Revision as of 11:10, 6 February 2019
Description
Abbreviation: N(PGM)
Reference: A »Versions
MitoPedia concepts:
MiP concept,
SUIT protocol,
Recommended
MitoPedia methods:
Respirometry
- SUIT-category: N(PGM)
- SUIT protocol pattern: diametral 1PM;2D;2c;3G;4U-
The SUIT-012 protocols specifically focus on assessing the linear coupling control (L- P- E) with NADH linked-substrates (PM) and the control in ET state (N). Addition of G in NADH-supported OXPHOS enables evaluating the glutamate anaplerotic pathway control state. SUIT-004 can be extended with the CIV assay module.
Communicated by Cardoso LH, Doerrier C, Huete-Ortega M, Iglesias-Gonzalez J and Gnaiger E (last update 2019-01-28)
Specific SUIT protocols
- SUIT-012 O2 mt D027 for isolated mitochondria
- SUIT-012 O2 pce D### for permeabilized cells
References
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
|
3G | PGMP | N | CI | 1PM;2D;2c;3G
|
4U | PGME | N | CI | 1PM;2D;2c;3G;4U
|
5Ama | ROX | 1PM;2D;2c;3G;4U;5Ama
|
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
- + This protocol allows to evaluate the function of the TCA cycle without the involvement of the complex II (S-pathway). It is thus useful to understand the contribution and the activity of dehydrogenases providing NADH.
- + Mitochondrial external membrane integrity can be measured with the addition of cytochrome c. Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.
- + Reasonable duration of the experiment.
- + Complex IV activity can be measured.
- + GM and PM yield typically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM: the fraction of the N-pathway is lower and of the S-pathway is higher with GM compared to PM (GMP is inhibited by the CII inhibitor malonic acid to a larger extent than PMP). PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-pathway, since an impairment of N-pathway capacity can be compensated partially by activation of the S-pathway. This is an advantage compared to SUIT-011 for diagnosis of N-capacity.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.