Flux control ratio: Difference between revisions
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For a given protocol or set of respiratory protocols, flux control ratios provide a fingerprint of coupling and substrate control independent of (i) mt-content in cells or tissues, (ii) purification in preparations of isolated mitochondria, and (iii) assay conditions for determination of tissue mass or mt-markers external to a respiratory protocol (CS, protein, stereology, etc.). ''FCR'' obtained from a single respirometric incubation with sequential titrations (sequential protocol; [[SUIT|SUIT protocol]]) provide an internal normalization, expressing respiratory control independent of mitochondrial content and thus independent of a marker for mitochondrial amount. ''FCR'' obtained from separate (parallel) protocols depend on equal distribution of subsamples obtained from a homogenous mt-preparation or determination of a common [[mitochondrial marker]]. | For a given protocol or set of respiratory protocols, flux control ratios provide a fingerprint of coupling and substrate control independent of (i) mt-content in cells or tissues, (ii) purification in preparations of isolated mitochondria, and (iii) assay conditions for determination of tissue mass or mt-markers external to a respiratory protocol (CS, protein, stereology, etc.). ''FCR'' obtained from a single respirometric incubation with sequential titrations (sequential protocol; [[SUIT|SUIT protocol]]) provide an internal normalization, expressing respiratory control independent of mitochondrial content and thus independent of a marker for mitochondrial amount. ''FCR'' obtained from separate (parallel) protocols depend on equal distribution of subsamples obtained from a homogenous mt-preparation or determination of a common [[mitochondrial marker]]. | ||
|info=[[Gnaiger 2014 MitoPathways]], [[Gnaiger 2009 Int J Biochem Cell Biol]], [[ | |info=[[Gnaiger 2014 MitoPathways]], [[Gnaiger 2009 Int J Biochem Cell Biol]], [[Doerrier 2018 Methods Mol Biol]] | ||
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== Flux control factor: normalization of mitochondrial respiration == | |||
::::» ''More details:'' [[Flux control factor]] | |||
== DatLab == | |||
=== Unknown sample concentration and normalization per unit sample [x] === | |||
:::* In new Excel Template for DatLab 7.4 | |||
:::: If the sample concentration is not yet known, the box ‘Known sample concentration’ can be unchecked, and the concentration will be considered by default as 1, with units [x·mL<sup>-1</sup>]. In this way, flux can be normalized and ''FCR''s can be obtained even if the sample concentration is unknown. [[Extensive_quantity]] | |||
:::: The entire plot of oxygen flux can be transformed to a ''FCR''. Click on 'Flux/Slope' in the pull-down menu. | |||
== References == | |||
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Revision as of 14:02, 17 May 2020
- high-resolution terminology - matching measurements at high-resolution
Flux control ratio
Description
Flux control ratios (FCR), are ratios of oxygen flux in different respiratory control states, normalized for maximum flux in a common reference state, to obtain theoretical lower and upper limits of 0.0 and 1.0 (0% and 100%).
For a given protocol or set of respiratory protocols, flux control ratios provide a fingerprint of coupling and substrate control independent of (i) mt-content in cells or tissues, (ii) purification in preparations of isolated mitochondria, and (iii) assay conditions for determination of tissue mass or mt-markers external to a respiratory protocol (CS, protein, stereology, etc.). FCR obtained from a single respirometric incubation with sequential titrations (sequential protocol; SUIT protocol) provide an internal normalization, expressing respiratory control independent of mitochondrial content and thus independent of a marker for mitochondrial amount. FCR obtained from separate (parallel) protocols depend on equal distribution of subsamples obtained from a homogenous mt-preparation or determination of a common mitochondrial marker.
Abbreviation: FCR
Reference: Gnaiger 2014 MitoPathways, Gnaiger 2009 Int J Biochem Cell Biol, Doerrier 2018 Methods Mol Biol
Flux control factor: normalization of mitochondrial respiration
- » More details: Flux control factor
DatLab
Unknown sample concentration and normalization per unit sample [x]
- In new Excel Template for DatLab 7.4
- If the sample concentration is not yet known, the box ‘Known sample concentration’ can be unchecked, and the concentration will be considered by default as 1, with units [x·mL-1]. In this way, flux can be normalized and FCRs can be obtained even if the sample concentration is unknown. Extensive_quantity
- The entire plot of oxygen flux can be transformed to a FCR. Click on 'Flux/Slope' in the pull-down menu.
References
| Bioblast link | Reference | Year |
|---|---|---|
| Doerrier 2018 Methods Mol Biol | 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 | 2018 |
| Gnaiger 2009 Int J Biochem Cell Biol | Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. https://doi.org/10.1016/j.biocel.2009.03.013 | 2009 |
| Gnaiger 2020 BEC MitoPathways | Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002 | 2020 |
| BEC 2020.1 doi10.26124bec2020-0001.v1 | Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1 | 2020 |
MitoPedia concepts:
Respiratory control ratio,
SUIT concept
MitoPedia methods:
Respirometry
MitoPedia O2k and high-resolution respirometry:
DatLab
