Flux control ratio

Flux control ratio

MitoPedia O2k and high-resolution respirometry: Oroboros Open Support

Description

Flux control ratios FCRs 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 (1) mt-content in cells or tissues, (2) purification in preparations of isolated mitochondria, and (3) 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 2020 BEC MitoPathways, Gnaiger 2009 Int J Biochem Cell Biol, Doerrier 2018 Methods Mol Biol

Flux control efficiency: normalization of mitochondrial respiration

» More details: Flux control efficiency

DatLab

Unknown sample concentration and normalization per unit sample [x]

In the DatLab 7.4 Excel template for oxygen flux analysis (O2 analysis template DL7.4):

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.

» Read also: Extensive quantity; BEC 2020.1

» More details: MiPNet24.06 Oxygen flux analysis - DatLab 7.4

FCR in DatLab plot

The entire oxygen flux plot can be converted to a FCR. Click on 'Flux/Slope' in the DatLab pull-down menu. Select chamber A or B 'O2 slope'. Select 'Flux control ratio, FCR' and select the mark that corresponds to the reference state. Change the layout under scale under 'Layout/Standard layouts' and select '07a Flux Control Ratios' or '07b Flux Control Ratios overlay'.

» More details: Flux control ratio

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. 5^th 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

Keywords

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The Blue Book 2020
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OXPHOS-, ROUTINE-, ET-, and LEAK states; respiratory capacities (P, R, E, L) corrected for residual oxygen consumption Rox.
4-compartmental OXPHOS model. (1) ET capacity E of the noncoupled electron transfer system ETS. OXPHOS capacity P is partitioned into (2) the dissipative LEAK component L, and (3) ADP-stimulated P-L net OXPHOS capacity. (4) If P-L is kinetically limited by a low capacity of the phosphorylation system to utilize the protonmotive force pmF, then the apparent E-P excess capacity is available to drive coupled processes other than phosphorylation P» (ADP to ATP) without competing with P».
(P-L)/P is the OXPHOS P-L control efficiency. The biochemical coupling efficiency is independent of kinetic control by the phosphorylation system when expressed as the E-L coupling efficiency, (E-L)/E. (E-P)/E is the kinetic E-P control efficiency.

Bioblast links: Coupling control - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

1. Mitochondrial and cellular respiratory rates in coupling-control states

» Baseline state

Respiratory rate	Defining relations
OXPHOS capacity	P = P´-Rox	mt-preparations
ROUTINE respiration	R = R´-Rox	living cells
ET capacity	E = E´-Rox	» Level flow
		» Noncoupled respiration - Uncoupler
LEAK respiration	L = L´-Rox	» Static head
		» LEAK state with ATP
		» LEAK state with oligomycin
		» LEAK state without adenylates
Residual oxygen consumption Rox	L = L´-Rox

Chance and Williams nomenclature: respiratory states

» State 1 —» State 2 —» State 3 —» State 4 —» State 5

2. Flux control ratios related to coupling in mt-preparations and living cells

» Flux control ratio

» Coupling-control ratio

» Coupling-control protocol

FCR	Definition
L/P coupling-control ratio	L/P	» Respiratory acceptor control ratio, RCR = P/L
L/R coupling-control ratio	L/R
L/E coupling-control ratio	L/E	» Uncoupling-control ratio, UCR = E/L (ambiguous)
P/E control ratio	P/E
R/E control ratio	R/E	» Uncoupling-control ratio, UCR = E/L
net P/E control ratio	(P-L)/E
net R/E control ratio	(R-L)/E

3. Net, excess, and reserve capacities of respiration

Respiratory net rate	Definition	Icon
P-L net OXPHOS capacity	P-L
R-L net ROUTINE capacity	R-L
E-L net ET capacity	E-L
E-P excess capacity	E-P
E-R reserve capacity	E-R

4. Flux control efficiencies related to coupling-control ratios

» Flux control efficiency j_Z-Y

» Background state

» Reference state

» Metabolic control variable

Coupling-control efficiency		Definition		Canonical term
P-L control efficiency	j_P-L	= (P-L)/P	= 1-L/P	P-L OXPHOS-flux control efficiency
R-L control efficiency	j_R-L	= (R-L)/R	= 1-L/R	R-L ROUTINE-flux control efficiency
E-L coupling efficiency	j_E-L	= (E-L)/E	= 1-L/E	E-L ET-coupling efficiency » Biochemical coupling efficiency
E-P control efficiency	j_E-P	= (E-P)/E	= 1-P/E	E-P ET-excess flux control efficiency
E-R control efficiency	j_E-R	= (E-R)/E	= 1-R/E	E-R ET-reserve flux control efficiency