Description
MitoPathway control state: NADH Electron transfer-pathway state
Upstream of the NAD-junction, Pyruvate (P) is oxidatively decarboxylated to acetyl-CoA and CO2, yielding NADH catalyzed by pyruvate dehydrogenase. Malate (M) is oxidized to oxaloacetate by mt-malate dehydrogenase located in the mitochondrial matrix. Condensation of oxaloacate with acetyl-CoA yields citrate (citrate synthase). 2-oxoglutarate (Ξ±-ketoglutarate) is formed from isocitrate (isocitrate dehydrogenase).
Abbreviation: PM
Reference: Gnaiger 2020 BEC MitoPathways
PML
With PM as N-substrates, LEAK respiration L can be evaluated in the following SUIT protocols:
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- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-001 O2 mt D001
- DL-Protocol for permeabilized fibers (pfi): SUIT-001 O2 pfi D002
- DL-Protocol for permeabilized cells (pce): SUIT-001 O2 ce-pce D003
- DL-Protocol for permeabilized blood cells (PBMC and PLT): SUIT-001 O2 ce-pce D004
- DL-Protocol for permeabilized fibers (pfi): SUIT-004 O2 pfi D010
- DL-Protocol for permeabilized cells (pce): SUIT-006 O2 ce-pce D029
- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-006 O2 mt D047
- DL-Protocol for permeabilized fibers (pfi): SUIT-008 O2 pfi D014
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PMP
With PM as N-substrates, OXPHOS capacity P can be evaluated in the following SUIT protocols:
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- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-001 O2 mt D001
- DL-Protocol for permeabilized fibers (pfi): SUIT-001 O2 pfi D002
- DL-Protocol for permeabilized cells (pce): SUIT-001 O2 ce-pce D003
- DL-Protocol for permeabilized blood cells (PBMC and PLT): SUIT-001 O2 ce-pce D004
- DL-Protocol for permeabilized fibers (pfi): SUIT-004 O2 pfi D010
- DL-Protocol for permeabilized cells (pce): SUIT-006 O2 ce-pce D029
- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-006 O2 mt D047
- DL-Protocol for permeabilized fibers (pfi): SUIT-008 O2 pfi D014
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PME
With PM as N-substrates, ET capacity E can be evaluated in the following SUIT protocols:
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- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-001 O2 mt D001
- DL-Protocol for permeabilized fibers (pfi): SUIT-001 O2 pfi D002
- DL-Protocol for permeabilized cells (pce): SUIT-001 O2 ce-pce D003
- DL-Protocol for permeabilized blood cells (PBMC and PLT): SUIT-001 O2 ce-pce D004
- DL-Protocol for permeabilized fibers (pfi): SUIT-004 O2 pfi D010
- DL-Protocol for permeabilized cells (pce): SUIT-006 O2 ce-pce D029
- DL-Protocol for isolated mitochondria and tissue homogenate (mt): SUIT-006 O2 mt D047
- DL-Protocol for permeabilized fibers (pfi): SUIT-008 O2 pfi D014
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Linear coupling control in the N-pathway control state: L β P β E
- P-L
- P-L control efficiency, jP-L = (P-L)/P = 1-L/P, is measured in the N-pathway state, with defined coupling sites (CI, CIII, CIV).
- P-E
- CCCP is titrated stepwise to maximum flux, to evaluate limitation of OXPHOS by the phosphorylation system, expressed as the E-P control efficiency jE-P = (E-P)/E = 1-P/E.
- If jE-P>0, then the E-L coupling efficiency rather than the P-L control efficiency is the proper expression of coupling, jE-L = (E-L)/E = 1-L/E.
Discussion
- The Pyruvate anaplerotic pathway control state (pyruvate alone) is not an ET-pathway competent substrate state in most mt-preparations, since acetyl-CoA accumulates without the co-substrate (oxaloacetate) of citrate synthase.
- The Malate-anaplerotic pathway control state (M alone) is not an ET-pathway competent substrate state in many mt-preparations, since oxaloacetate accumulates without the co-substrate (acetyl-CoA) of citrate synthase.
MitoPedia concepts:
SUIT state