SUIT-003 O2 ce D009

high-resolution terminology - matching measurements at high-resolution

SUIT-003 O2 ce D009

Description

Reference: A Huetter 2004 Biochem J

SUIT number: 12

MitoPedia concepts: SUIT protocol, SUIT A 

SUIT protocol pattern: linear cell (ce) coupling control protocol (ceCCP)

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 (4ceRot) and antimycin A (5ceAma). In many intact cells ROX measured after titration of rotenone (4ceRot) is not or only slightly further inhibited by subsequent titration of animycin A (5ceAma).

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.

• 1ce: ROUTINE (R; routine state in cell-culture medium) in intact cells (ce);
• 2ceOmy: LEAK (L; inhibition of ATPsynthase by 1 μg/ml oligomycin);
• 3ceU: ET-pathway (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration);
• 4ceRot: ROX (rotenone, Rot; residual oxygen consumption).
• 5ceAma: ROX (antimycin A, Ama; 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«