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Difference between revisions of "SUIT-003 O2 ce D061"

From Bioblast
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|abbr=CCP-ce Snv,Mnanv - control
|abbr=CCP-ce Snv,Mnanv - control
|description=[[File:ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama.png|400px]]
|description=[[File:ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama.png|400px]]
|info='''B''' Carrier control for the [[coupling control protocol]] for [[living cells]] with cell permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]] - [[SUIT-003 O2 ce D060]]
|info='''B''' Carrier control for the [[coupling-control protocol]] for [[living cells]] with cell permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]] - [[SUIT-003 O2 ce D060]]
|application=O2
|application=O2
|SUIT number=D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
|SUIT number=D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
}}
}}
::: '''[[MitoPedia: SUIT]]''' - Coupling control protocol (CCP) with the carrier of the plasma membrane-permeable succinate ([[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate ([[MitoKit-CII/Malonate-nv]]) in living cells (ce): DMSO.
::: '''[[MitoPedia: SUIT]]''' - Coupling-control protocol (CCP) with the carrier of the plasma membrane-permeable succinate ([[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate ([[MitoKit-CII/Malonate-nv]]) in living cells (ce): DMSO.
::: '''[[SUIT protocol pattern]]:''' ce1;(ce2Omy);ce3U;ce4DMSO;ce5DMSO;ce6Rot;ce7Ama
::: '''[[SUIT protocol pattern]]:''' ce1;(ce2Omy);ce3U;ce4DMSO;ce5DMSO;ce6Rot;ce7Ama


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::::* [[SUIT-003 O2 ce D060]] CCP with plasma membrane-permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]].
::::* [[SUIT-003 O2 ce D060]] CCP with plasma membrane-permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]].
::::* [[SUIT-003 O2 ce D028]] for cell membrane permeability test CCP(S).
::::* [[SUIT-003 O2 ce D028]] for cell membrane permeability test CCP(S).
::::* [[SUIT-003 O2 ce-pce D013]] first version of the Coupling control protocol and respirometric cell viability test (CCV protocol).
::::* [[SUIT-003 O2 ce-pce D013]] first version of the Coupling-control protocol and respirometric cell viability test (CCV protocol).
::::* [[SUIT-003 O2 ce-pce D018]] second version of the Coupling control protocol and respirometric cell viability test (CCV protocol).
::::* [[SUIT-003 O2 ce-pce D018]] second version of the Coupling-control protocol and respirometric cell viability test (CCV protocol).
::::* [[SUIT-003 Ce1;ce1P;ce3U;ce4Glc;ce5M;ce6Rot;ce7S;1Dig;1c;2Ama;3AsTm;4Azd]] coupling control protocol and respirometric cell viability test (CCV protocol) without oligomycin.
::::* [[SUIT-003 Ce1;ce1P;ce3U;ce4Glc;ce5M;ce6Rot;ce7S;1Dig;1c;2Ama;3AsTm;4Azd]] coupling-control protocol and respirometric cell viability test (CCV protocol) without oligomycin.
::::* [[SUIT-003 Ce1;ce2U-]]
::::* [[SUIT-003 Ce1;ce2U-]]
::::* [[SUIT-003 Ce1;ce2SD;ce3Omy;ce4U-]] Β 
::::* [[SUIT-003 Ce1;ce2SD;ce3Omy;ce4U-]] Β 
::::* [[SUIT-003 Ce1;ce2SD;ce3U;ce4Rot;ce5Ama]]
::::* [[SUIT-003 Ce1;ce2SD;ce3U;ce4Rot;ce5Ama]]
::::* [[SUIT-003 Ce1;ce2U;ce3Rot;ce4S;ce5Ama]]
::::* [[SUIT-003 Ce1;ce2U;ce3Rot;ce4S;ce5Ama]]
::::* [[SUIT-006]]: Coupling control protocol for mt-preparations (mtprep), counterpart to SUIT-003 for non-permeabilized cells (ce).
::::* [[SUIT-006]]: Coupling-control protocol for mt-preparations (mtprep), counterpart to SUIT-003 for non-permeabilized cells (ce).





Revision as of 15:26, 2 June 2020


high-resolution terminology - matching measurements at high-resolution


SUIT-003 O2 ce D061

Description

Ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama.png

Abbreviation: CCP-ce Snv,Mnanv - control

Reference: B Carrier control for the coupling-control protocol for living cells with cell permeable compounds MitoKit-CII/Succinate-nv and MitoKit-CII/Malonate-nv - SUIT-003 O2 ce D060

SUIT number: D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama

O2k-Application: O2

MitoPedia: SUIT - Coupling-control protocol (CCP) with the carrier of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv) in living cells (ce): DMSO.
SUIT protocol pattern: ce1;(ce2Omy);ce3U;ce4DMSO;ce5DMSO;ce6Rot;ce7Ama

SUIT-003 O2 ce D061 protocol is a control for the SUIT-003 O2 ce D060, with the addition of the carrier DMSO instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv). Moreover, the SUIT-003 O2 ce D061 protocol allows evaluating the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and Electron transfer pathway. Optionally, to study LEAK respiration, the phosphorylation system might be inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET capacity. SUIT-003 O2 ce D061 protocol is a control for the SUIT-003 O2 ce D060, with the addition of the carrier DMSO instead of the plasma membrane-permeable succinate ([[[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv). Moreover, the SUIT-003 O2 ce D061 protocol allows evaluating the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and ET. Optionally, to study LEAK-respiration, the phosphorylation system might be inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET-capacity.


Communicated by  Garcia-Souza LF, Cardoso LHD and Gnaiger E(last update 2019-07-02)
MitoPedia: SUIT

Steps and respiratory states

Ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama.png

Step State Pathway Q-junction 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.


(ce2Omy) L(Omy) ce1;(ce2Omy)
  • Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
  • The addition of oligomycin is optional depending on the experimental question to resolve.
ce3U E cce1;(ce2Omy);ce3U
ce4Rot ROX ce1;(ce2Omy);ce3U;ce4Rot
  • 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.
ce5DMSO ROX ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO
  • 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.
ce6DMSO ROX ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO
  • 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.
ce7Ama ROX ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
  • 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.

Strengths and limitations

+ This protocol is a control for SUIT-003 O2 ce D060, with titrations of the carrier DMSO.
+ Reasonable duration of the experiment.
- To obtain LEAK respiration, the concentration of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET-capacity.
- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.


Compare SUIT protocols


References

MitoPedia concepts: SUIT protocol, SUIT B, Find 


MitoPedia methods: Respirometry