SUIT-003 O2 ce D009: Difference between revisions
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' RLE | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' RLE | ||
::: '''[[SUIT protocol pattern]]:''' linear [[coupling control protocol]] ( | ::: '''[[SUIT protocol pattern]]:''' linear cell (ce) [[coupling control protocol]] (ceCCP) | ||
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== R;L;E format_ROX == | == R;L;E format_ROX == | ||
[[File:CCP_02(R,L,E).jpg|left|150px]] | [[File:CCP_02(R,L,E).jpg|left|150px]] | ||
:::* The | :::* The ceCCP diagram may show the respiratory states directly (left) as an alternative to presenting the titration steps. | ||
:::* '''Rot and Ama''' Β | :::* '''Rot and Ama''' Β | ||
:::: In this ceCCP, ROX is measured after titration of [[rotenone]], [[antimycin A]], or both in a single step. Validity of using [[rotenone]] only for inducing ROX in intact cells is evaluated by stepwise measurement of ROX after titration of rotenone (3Rot) followed by titration of animycin A (4ceAma). | :::: In this ceCCP, ROX is measured after titration of [[rotenone]], [[antimycin A]], or both in a single step. Validity of using [[rotenone]] only for inducing ROX in intact cells is evaluated by stepwise measurement of ROX after titration of rotenone (3Rot) followed by titration of animycin A (4ceAma). | ||
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:::: In this ceCCP, ROX is measured after stepwise titration of [[rotenone]] (3ceRot) and [[antimycin A]] (4ceAma). In many intact cells ROX measured after titration of rotenone (3ceRot) is not or only slightly further inhibited by subsequent titration of animycin A (4ceAma). | :::: In this ceCCP, ROX is measured after stepwise titration of [[rotenone]] (3ceRot) and [[antimycin A]] (4ceAma). In many intact cells ROX measured after titration of rotenone (3ceRot) is not or only slightly further inhibited by subsequent titration of animycin A (4ceAma). | ||
[[File:Huetter 2004 Biochem J.jpg|left|300px| | [[File:Huetter 2004 Biochem J.jpg|left|300px|ceCCP (fibroblasts)]] | ||
<br /> | <br /> | ||
:::: Respiration in senescent human primary fibroblasts (0.2Γ10<sup>6</sup> cells/ml). Arrows show steps in the titration regime of the [[coupling control protocol]], inducing the following respiratory states. Β | :::: Respiration in senescent human primary fibroblasts (0.2Γ10<sup>6</sup> cells/ml). Arrows show steps in the titration regime of the [[coupling control protocol]], inducing the following respiratory states. Β | ||
::::* 1ce: ROUTINE (''R''; routine state in cell-culture medium); Β | ::::* 1ce: ROUTINE (''R''; routine state in cell-culture medium) in intact cells (ce); Β | ||
::::* 2ceOmy: LEAK (''L''; inhibition of ATPsynthase by 1 ΞΌg/ml oligomycin); Β | ::::* 2ceOmy: LEAK (''L''; inhibition of ATPsynthase by 1 ΞΌg/ml oligomycin); Β | ||
::::* 3ceU: ETS (''E''; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5β4 ΞΌM final concentration); Β | ::::* 3ceU: ETS (''E''; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5β4 ΞΌM final concentration); Β |
Revision as of 08:20, 5 August 2017
Description
Abbreviation: RLE
Reference: A Huetter 2004 Biochem J
MitoPedia concepts:
SUIT protocol,
SUIT A
- SUIT-category: RLE
- SUIT protocol pattern: linear cell (ce) coupling control protocol (ceCCP)
References
R;L;E format_ROX
- The ceCCP diagram may show the respiratory states directly (left) as an alternative to presenting the titration steps.
- Rot and Ama
- In this ceCCP, ROX is measured after titration of rotenone, antimycin A, or both in a single step. Validity of using rotenone only for inducing ROX in intact cells is evaluated by stepwise measurement of ROX after titration of rotenone (3Rot) followed by titration of animycin A (4ceAma).
1ce;2ceOmy;3ceU;4ceRot;5ceAma
- 1ce;2ceOmy;3ceU;4ceRot;5ceAma
- In this ceCCP, ROX is measured after stepwise titration of rotenone (3ceRot) and antimycin A (4ceAma). In many intact cells ROX measured after titration of rotenone (3ceRot) is not or only slightly further inhibited by subsequent titration of animycin A (4ceAma).
- Respiration in senescent human primary fibroblasts (0.2Γ106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- 1ce: ROUTINE (R; routine state in cell-culture medium) in intact cells (ce);
- 2ceOmy: LEAK (L; inhibition of ATPsynthase by 1 ΞΌg/ml oligomycin);
- 3ceU: ETS (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5β4 ΞΌM final concentration);
- 4ceRot: ROX (rotenone, Rot; residual oxygen consumption).
- 5ceAma: ROX (antimycin A, Ama; residual oxygen consumption). Modified after Huetter_2004_Biochem J.
- Respiration in senescent human primary fibroblasts (0.2Γ106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- HΓΌtter E, Renner K, Pfister G, StΓΆckl P, Jansen-DΓΌrr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. - Β»Bioblast linkΒ«