Difference between revisions of "MiPNet12.01 Suppl T-issue"
From Bioblast
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{{OROBOROS header page name}} | |||
{{Publication | {{Publication | ||
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols contents]] MitoPathways at the Q-junction: mouse skeletal muscle fibers. | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols contents]] MitoPathways at the Q-junction: mouse skeletal muscle fibers. | ||
|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/2/2d/MiPNet12.01_Suppl_T-issue.pdf |Bioblast pdf]] »[http://www.bioblast.at/index.php/File:MiPNet12.01_Suppl_T-issue.pdf Versions] | |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/2/2d/MiPNet12.01_Suppl_T-issue.pdf |Bioblast pdf]] »[http://www.bioblast.at/index.php/File:MiPNet12.01_Suppl_T-issue.pdf Versions] | ||
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|year=2018-11-18 | |year=2018-11-18 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
|abstract= | |abstract= Oroboros (2018) MitoPathways at the Q-junction: mouse skeletal muscle fibers. Mitochondr Physiol Network 12.01(03): Suppl T-issue. » [http://www.bioblast.at/index.php/File:MiPNet12.01 Versions] | ||
{{MiPNet pdf page linking to MitoPedia}} | |||
|mipnetlab=AT_Innsbruck_Oroboros | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
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|additional=MitoPathways, O2k-Demo, O2k-Core | |additional=MitoPathways, O2k-Demo, O2k-Core | ||
}} | }} | ||
== Limitations of the SUIT protocol == | == Limitations of the SUIT protocol == | ||
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=== ROX correction === | === ROX correction === | ||
:::: The fact that ROX was higher in the NS-substrate state compared to N-linked LEAK respiration indicates that ROX is partially controlled by the substrate state. Therefore, a single measurement of ROX cannot be applied for correction of total oxygen consumption in the different substrate states. Total respiration, therefore, represents apparent coupling states ''L''´, ''P''´ and ''E''´ (Fig. 1). ROX correction is possible in the present experiment only for NS- and S-linked respiration. [[Azide]] inhibits not only CIV but other heme-based oxidases and peroxidases, and therefore may interfere with ROX beyond blocking respiratory electron transfer. Based on this argument, a combination of CII- and CIII-inhibitors (malonic acid, antimycin A, myxothiazol) may yield more consistent results, although any ROS scavenged by cytochrome ''c'' may in the absence of a CIV-inhibitor result in respiratory oxygen consumption through CIV. | :::: The fact that ROX was higher in the NS-substrate state compared to N-linked LEAK respiration indicates that ROX is partially controlled by the substrate state. Therefore, a single measurement of ROX cannot be applied for correction of total oxygen consumption in the different substrate states. Total respiration, therefore, represents apparent coupling states ''L''´, ''P''´ and ''E''´ (Fig. 1). ROX correction is possible in the present experiment only for NS- and S-linked respiration. [[Azide]] inhibits not only CIV but other heme-based oxidases and peroxidases, and therefore may interfere with ROX beyond blocking respiratory electron transfer. Based on this argument, a combination of CII- and CIII-inhibitors (malonic acid, antimycin A, myxothiazol) may yield more consistent results, although any ROS scavenged by cytochrome ''c'' may in the absence of a CIV-inhibitor result in respiratory oxygen consumption through CIV. | ||
Revision as of 12:29, 4 December 2019
MiPNet12.01 Suppl T-issue
MitoPathways at the Q-junction: mouse skeletal muscle fibers. |
» »Versions
Oroboros (2018-11-18) Mitochondr Physiol Network
Abstract: Oroboros (2018) MitoPathways at the Q-junction: mouse skeletal muscle fibers. Mitochondr Physiol Network 12.01(03): Suppl T-issue. » Versions
- » Current O2k-series: NextGen-O2k Series XB and O2k Series J
- » Current software versions DatLab 8.0: MitoPedia: DatLab
- Further details: » MitoPedia: O2k-Open Support
• O2k-Network Lab: AT_Innsbruck_Oroboros
Labels: MiParea: Respiration
Organism: Mouse
Tissue;cell: Skeletal muscle
Preparation: Permeabilized tissue
Coupling state: LEAK, ROUTINE, ET
Pathway: N, S, NS, ROX
HRR: Oxygraph-2k, O2k-Protocol
MitoPathways, O2k-Demo, O2k-Core
Limitations of the SUIT protocol
Maximum OXPHOS and ET-capacity
- Evaluation of maximum respiratory capacities requires titration of further substrates activating additional respiratory complexes at the Q-junction (CETF and CGpDH<).
Malate concentration
- The malate concentration was 2 mM, to saturate N-linked respiration. However, at 2 mM malate, the fumarate concentration is increased to a level which inhibits succinate dehydrogenase. Then NS- and S-linked respiratory capacities are underestimated. A malate concentration of 0.5 mM, which saturates N-linked respiration and inhibits S-linked respiration to a lesser extent, represents and improved standard.
- » Optimum malate concentration in SUIT protocols
ROX correction
- The fact that ROX was higher in the NS-substrate state compared to N-linked LEAK respiration indicates that ROX is partially controlled by the substrate state. Therefore, a single measurement of ROX cannot be applied for correction of total oxygen consumption in the different substrate states. Total respiration, therefore, represents apparent coupling states L´, P´ and E´ (Fig. 1). ROX correction is possible in the present experiment only for NS- and S-linked respiration. Azide inhibits not only CIV but other heme-based oxidases and peroxidases, and therefore may interfere with ROX beyond blocking respiratory electron transfer. Based on this argument, a combination of CII- and CIII-inhibitors (malonic acid, antimycin A, myxothiazol) may yield more consistent results, although any ROS scavenged by cytochrome c may in the absence of a CIV-inhibitor result in respiratory oxygen consumption through CIV.