SUIT-010 O2 ce-pce D008

high-resolution terminology - matching measurements at high-resolution

SUIT-010 O2 ce-pce D008

Description

Abbreviation: ceS,S

Reference: A SUIT-010 Optimum digitonin concentration for pce  »Versions

SUIT number: D008_ce1;ce2Rot;ce3S;ce4D;1Dig;1c

O2k-Application: O2

SUIT-category: ceS,S

SUIT protocol pattern: diametral ce1;ce2Rot;ce3S;ce4D;1Dig;1c

Communicated by Huete-Ortega M and Gnaiger E (last update 2019-01-22)

Steps and respiratory states

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.
ce2Rot	ROX			ce1;ce2Rot Succinate pathway control state (S-pathway) after inhibiting CI with rotenone, which also inhibits the F-pathway.
ce3S	SL(n)	S	CII	ce1;ce2Rot;ce3S Succinate, S ( type S-pathway to Q). Non-phosphorylating resting state (LEAK state); LEAK respiration L(n) in the absence of ADP, ATP, AMP (no adenylates).
ce4D	SP	S	CII	ce1;ce2Rot;ce3S;ce4D Succinate, S ( type S-pathway to Q). OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
1Dig	SP	S	CII	ce1;ce2Rot;ce3S;ce4D;1Dig Succinate, S ( type S-pathway to Q). Multiple Digitonin titrations for complete plasma membrane permeabilization. OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
1c	SP	S	CII	ce1;ce2Rot;ce3S;ce4D;1Dig;1c Succinate, S ( type S-pathway to Q). Addition of cytochrome c yields a test for integrity of the mtOM (cytochrome c control efficiency). Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (OXPHOS capacity P with saturating [ADP]). OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.

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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

• Digitonin is a natural product and thus the effective concentration has to be determined by titrations for every batch. The optimum effective digitonin concentrations for complete plasma membrane permeabilization can be determined with this protocol.

Compare SUIT protocols

• SUIT-003 have been designed to study the coupling control state of intact cells
• SUIT-003 O2 ce D028 coupling control protocol with succinate for permeability test
• SUIT-006 protocol is a coupling control protocol for mitochondrial preparations

References

MitoPedia concepts: SUIT protocol, SUIT A, Find 

MitoPedia methods: Respirometry 

Labels:

SUIT-010