SUIT-003 O2 ce D009: Difference between revisions

Revision as of 12:18, 3 September 2016

high-resolution terminology - matching measurements at high-resolution

SUIT-003 O2 ce D009

Description

File:CCP R 1Omy 2U.jpg

Abbreviation: CCP_R 1Omy 2U

Reference: MiPNet08.09 CellRespiration, Huetter 2004 Biochem J

MitoPedia concepts: SUIT protocol

SUIT-catg: CCP

SUIT protocol pattern: linear coupling control protocol (CCP)

Mark names in DatLab: CCP_01

DatLab-Excel template: CCP_01.xlsx

File:CCP R-L-E-ROX.jpg

The CCP diagram may present the respiratory states (left) as an alternative to presenting the titration steps.
Rot and Ama

In this CCP, 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 (4Ama).

File:CCP R 1Omy 2U 3Rot 4Ama.jpg

CCP_01a

CCP_R 1Omy 2U 3Rot 4Ama
Rot and Ama

In this CCP, ROX is measured after stepwise titration of rotenone (3Rot) and antimycin A (4Ama). In many intact cells ROX measured after titration of rotenone (3Rot) is not or only slightly further inhibited by subsequent titration of animycin A (4Ama).

» Compare: CCP_01, Ce_R 1Omy 2U 3Rot

Respiration in senescent human primary fibroblasts (0.2×10⁶ cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states: ROUTINE (R; routine state in cell-culture medium); LEAK (L; inhibition of ATPsynthase by 1 μg/ml oligomycin); ETS (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration); ROX (rotenone, Rot, and antimycin A; residual oxygen consumption). Modified after Huetter_2004_Biochem J.

Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. - »Bioblast link«

@@ Line 1: / Line 1: @@
 {{MitoPedia
-|abbr=CC_R 1Omy 2U 3Ama
+|abbr=CCP_R 1Omy 2U
 |description=[[File:CCP R 1Omy 2U.jpg|right|150px]]
-|info=[[MiPNet08.09 CellRespiration]]
+|info=[[MiPNet08.09 CellRespiration]], [[Huetter 2004 Biochem J]]
 }}
 {{MitoPedia concepts
@@ Line 11: / Line 11: @@
 ::: '''[[Mark names in DatLab]]:''' CCP_01
 ::: '''[[DatLab-Excel templates |DatLab-Excel template]]:''' [[Media:SUIT MiPNet08.09 CellRespiration.xlsx|CCP_01.xlsx]]
+[[File:CCP R-L-E-ROX.jpg|left|150px]]
+:::* The CCP diagram may present the respiratory states (left) as an alternative to presenting the titration steps.
 :::* '''Rot and Ama'''
 :::: In this CCP, 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 (4Ama).
+[[File:CCP_R 1Omy 2U 3Rot 4Ama.jpg|right|200px]]
 == CCP_01a ==
-::::* CCP_R 1Omy 2U 3Rot 4Ama
+:::* CCP_R 1Omy 2U 3Rot 4Ama
+:::* '''Rot and Ama'''
+:::: In this CCP, ROX is measured after stepwise titration of [[rotenone]] (3Rot) and [[antimycin A]] (4Ama). In many intact cells ROX measured after titration of rotenone (3Rot) is not or only slightly further inhibited by subsequent titration of animycin A (4Ama).
+::::» ''Compare'': [[CCP_01]], Ce_R 1Omy 2U 3Rot
+[[File:Huetter 2004 Biochem J.jpg|left|300px|CCP intact cells (fibroblasts)]]
+<br />
+:::: Respiration in senescent human primary fibroblasts (0.2×10<sup>6</sup> cells/ml). Arrows show steps in the titration regime of the [[coupling control protocol]], inducing the following respiratory states: ROUTINE (''R''; routine state in cell-culture medium); LEAK (''L''; inhibition of ATPsynthase by 1 μg/ml oligomycin); ETS (''E''; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration); ROX (rotenone, Rot, and antimycin A; residual oxygen consumption). Modified after [[Huetter_2004_Biochem J]].
+:::* Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. - [[Huetter_2004_Biochem J |»Bioblast link«]]