SUIT-022 O2 ce D051

high-resolution terminology - matching measurements at high-resolution

SUIT-022 O2 ce D051

Description

Abbreviation: ?

Reference: A Determination of the respiration due to the alternative oxidase pathway -SUIT-022

SUIT number: D051_ce1;ce2KCN;ce3SHAM

O2k-Application: O2

MitoPedia: SUIT

SUIT-category: ?

SUIT protocol pattern: ? ce1;ce2KCN;ce3SHAM

The SUIT-022 O2 ce D051 has been designed to provide an estimation of the cyanide-and antimycin A-resistant mitochondrial respiration due to the activity of the alternative oxidase on cells on ROUTINE. To obtain a complete estimation, this protocol has to be carried out in parallel to SUIT-023 O2 ce D053

Communicated by Iglesias-Gonzalez J, Huete-Ortega M and Gnaiger E (last update 2019-04-09)

Steps and respiratory states

Step	State	Comment - Events (E) and Marks (M)
ce1	ROUTINE	ce1 ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
ce2KCN	ROX or ROUTINE	ce1;ce2KCN Cyanide (usually added as KCN) is a competitive inhibitor of cytochrome c oxidase (CIV). Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated either after inhibition of CIII (e.g. antimycin A, myxothiazol), CIV (e.g. Cyanide) or in the absence of endogenous fuel-substrates. Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration.
ce3SHAM	ROX	ce1;ce2KCN;ce3SHAM Salicylhydroxamic acid (SHAM) is an inhibitor of the alternative oxidase (AOX). When AOX is blocked by SHAM, electrons are forced through the cytochrome c oxidase (CIV) pathway. Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated either after inhibition of CIII (e.g. antimycin A, myxothiazol), CIV (e.g. Cyanide) or in the absence of endogenous fuel-substrates. Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration.

Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Coupling control

» Coupling control state

» ET capacity

» OXPHOS capacity

» LEAK respiration

Pathway control

» Electron transfer pathway

» Fatty acid oxidation pathway control state, F

» NADH electron transfer-pathway state, N

» Succinate pathway control state, S

» NS-pathway control state, NS

» Glycerophosphate pathway control state, Gp

» Complex IV single step, CIV

» Anaplerotic pathway control state

Main fuel substrates

» Glutamate, G

» Glycerophosphate, Gp

» Malate, M

» Octanoylcarnitine, Oct

» Pyruvate, P

» Succinate, S

Glossary

» List of SUIT states

» SUIT concept

Strengths and limitations

SUIT-022 O2 ce D051 provides an estimation of the cyanide-and antimycin A-resistant mitochondrial respiration due to the activity of the alternative oxidase on cells on ROUTINE

+ This protocol can be used with many type of cells, although optimal concentrations of inhibitors have to be optimised.

+ Short duration of experiment

- Estimation of the linear coupling control is not done

- The use of cyanide requires a careful and in deep cleaning of the chambers after use and careful handling of the chemical to guarantee health safety.

Compare SUIT protocols

SUIT-023 O2 ce D053 has to be used in parallel to obtain complete information of the alternative oxidase pathway contribution to mitochondrial respiration.

References

MitoPedia concepts: SUIT protocol, SUIT A, Find

MitoPedia methods: Respirometry