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Difference between revisions of "SUIT-003 O2 ce D060"

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ย  Communicated byย  [[Garcia-Souza LF]] (last update 2019-05-20)
ย  Communicated byย  [[Garcia-Souza LF]] (last update 2019-05-21)




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== Strengths and limitations ==
== Strengths and limitations ==

Revision as of 11:51, 21 May 2019


high-resolution terminology - matching measurements at high-resolution


SUIT-003 O2 ce D060

Description

Ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv;ce7Ama.png

Abbreviation: ce

Reference: B - SUIT-003 coupling control protocol (CCP) with cell permeable compounds NV118 and NV161

SUIT number: D060_ce1;ce2Omy;ce3U;ce4Rot;ce5Snv;ce6Mnanv;ce7Ama

O2k-Application: O2

SUIT-category: ce
SUIT protocol pattern: complex cell coupling control protocol ce1;ce2Omy;ce3U;ce*


Communicated by  Garcia-Souza LF (last update 2019-05-21)


References

MitoPedia: SUIT

Steps and respiratory states

Ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv;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 ce1;(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.


ce5Snv E ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv
  • Noncoupled electron transfer state, ET state, with ET capacity E.
  • MitoKit-CII/Succinate-nv (diacetoxymethyl succinate) is a plasma membrane-permeable prodrug (permeable succinate; Snv) that diffuses across the plasma membrane. Cleavage of diacetoxymethyl residues is mediated by intracellular esterases, thus releasing succinate in the intracellular space. Abliva #: 01-118-s4
ce6Mnanv ROX ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv
  • 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.
  • MitoKit-CII/Malonate-nv (diacetoxymethyl malonate) is a plasma membrane-permeable prodrug (permeable malonate; Mnanv) that diffuses across the plasma membrane. Cleavage of diacetoxymethyl residues is mediated by intracellular esterases, thus releasing malonate in the intracellular space. Abliva #: 01-161-s2
ce7Ama ROX ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv;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

Work in progress
- The addition 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