SUIT-011: Difference between revisions

Revision as of 10:10, 30 August 2016

high-resolution terminology - matching measurements at high-resolution

SUIT-011

Description

File:NS 1GM 2D 3c 4S 5U 6Rot 7Ama.png

SUIT-catg: NS

Abbreviation: NS_1GM 2D 3c 4S 5U 6Rot 7Ama

Reference: MiPNet12.23 FibreRespiration

MitoPedia concepts: SUIT protocol

Subcatg: N+NS+S

SUIT protocol pattern: diametral

Mark names in DatLab: MiPNet12_23_FibreRespiration

DatLab-Excel template: SUIT_MiPNet12.23_FibreRespiration.xlsx

SUIT states: 1-3GM(LPc) 4-5GMS(PE) 6S 7ROX

Step	Respiratory state	Pathway control	Pathway to Q	Comment
1GM	GM(L)	N	CI	LEAK state with type N substrates, N_L: Non-phosphorylating resting state with NADH-linked (type N) substrates glutamate&malate (GM; without adenalytes; CI-linked pathway to Q). See 2D.
2D	GM(P)	N	CI	OXPHOS capacity with type N substrates, NP: Respiratory capacity in the active coupled state (GM with ADP). GM and PM yield practically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM, since the fractions of the N-pathway is lower and of the S-pathway is higher with GM compared to PM. PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-linked (CI-linked) pathway. Compare SUIT targetRP1 NS and SUIT RP1.
3c	GM(c)	N	CI	Cytochrome c test for quality control of the integrity of the outer mitochondrial membrane (loss of cytochrome c is indicated by a stimulation of respiration). Cytochrome c added immediately after the earliest ADP-activation step. Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c (Gnaiger 2007 MitoPathways).
3S	GMS(P)	NS	CI&II	OXPHOS capacity with type NS substrates (CI&II-linked pathway to Q), NS_P: Respiratory stimulation by convergent electron flow through Complexes I&II at the Q-junction, in the coupled state after further addition of succinate (S), as an estimate of OXPHOS capacity with reconstitution of the TCA cycle (Gnaiger 2009 Int J Biochem Cell Biol).
5U	GMS(E)	NS	CI&II	Electron transfer system (ETS) capacity with type NS substrates, NS_E: Uncoupling by CCP or FCCP titration (avoiding inhibition by high uncoupler concentrations), as a test for limitation of OXPHOS relative to ETS capacity by the phosphorylation system. Limitation: The additive effect of N+S measured separately compared to the combined NS pathway cannot be evaluated in the same coupling state. With N_P, NS_P, NS_E, and S_E. If independent information is available on S_P = S_E, the additivity can be calculated for the OXPHOS state.
6Rot	S(E)	S	CII	ETS capacity with type S (CII) substrate, S_E: ETS capacity with succinate, after blocking Complex I with rotenone. Limitation: A succinate concentration of >10 mM may be required for saturating S_E capacity.
10Ama		ROX	ROX	Residual oxygen consumption (ROX) due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of Complex III). ROX may be lower in substrate states earlier in the SUIT protocol. Therefore, this ROX measurement is frequently taken as a methodological control rather than as the final basis of ROX correction of mitochondrial respiration (mt).

@@ Line 12: / Line 12: @@
 ::: '''[[Mark names in DatLab]]:''' MiPNet12_23_FibreRespiration
 ::: '''[[DatLab-Excel templates |DatLab-Excel template]]:''' SUIT_MiPNet12.23_FibreRespiration.xlsx
+<br />
 ::: '''SUIT states:''' 1-3[[GM]](LPc) 4-5[[GMS]](PE) 6[[S]] 7[[ROX]]
@@ Line 28: / Line 30: @@
 | [[N]]
 | CI
-| ''See'' 2D.
+| LEAK state with type N substrates, N<sub>''L''</sub>: Non-phosphorylating resting state with NADH-linked (type N) substrates glutamate&malate (GM; without adenalytes; CI-linked pathway to Q). ''See'' 2D.
@@ Line 36: / Line 38: @@
 | [[N]]
 | CI
-| GM and PM yield practically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM, since the fractions of the N-pathway is lower and of the S-pathway is higher with GM compared to PM. PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-linked (CI-linked) pathway. ''Compare'' [[SUIT targetRP1 NS]] and [[SUIT RP1]].
+| OXPHOS capacity with type N substrates, NP: Respiratory capacity in the active coupled state (GM with ADP). GM and PM yield practically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM, since the fractions of the N-pathway is lower and of the S-pathway is higher with GM compared to PM. PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-linked (CI-linked) pathway. ''Compare'' [[SUIT targetRP1 NS]] and [[SUIT RP1]].
 |-
@@ Line 43: / Line 46: @@
 | [[N]]
 | CI
-| Cytochrome ''c'' added immediately after the earliest ADP-activation step. Application of the cytochrome ''c'' test early in the protocol ensures comparability of all states in case of any effect of ''c'' ([[Gnaiger 2007 MitoPathways]]).
+| Cytochrome c test for quality control of the integrity of the outer mitochondrial membrane (loss of cytochrome c is indicated by a stimulation of respiration). Cytochrome ''c'' added immediately after the earliest ADP-activation step. Application of the cytochrome ''c'' test early in the protocol ensures comparability of all states in case of any effect of ''c'' ([[Gnaiger 2007 MitoPathways]]).
 |-
@@ Line 50: / Line 54: @@
 | [[NS]]
 | CI&II
-|
+| OXPHOS capacity with type NS substrates (CI<small>&</small>II-linked pathway to Q), NS<sub>''P''</sub>: Respiratory stimulation by convergent electron flow through Complexes I<small>&</small>II at the Q-junction, in the coupled state after further addition of succinate (S), as an estimate of OXPHOS capacity with reconstitution of the TCA cycle ([[Gnaiger 2009 Int J Biochem Cell Biol]]).
 |-
@@ Line 57: / Line 61: @@
 | [[NS]]
 | CI&II
-| The additive effect of N+S measured separately compared to the combined NS pathway cannot be evaluated in the same coupling state. With N<sub>''P''</sub>, NS<sub>''P''</sub>, NS<sub>''E''</sub>, and S<sub>''E''</sub>. If  independent information is available on S<sub>''P''</sub> =  S<sub>''E''</sub>, the additivity can be calculated for the OXPHOS state.
+| Electron transfer system (ETS) capacity with type NS substrates, NS<sub>''E''</sub>: Uncoupling by CCP or FCCP titration (avoiding inhibition by high uncoupler concentrations), as a test for limitation of OXPHOS relative to ETS capacity by the phosphorylation system. Limitation: The additive effect of N+S measured separately compared to the combined NS pathway cannot be evaluated in the same coupling state. With N<sub>''P''</sub>, NS<sub>''P''</sub>, NS<sub>''E''</sub>, and S<sub>''E''</sub>. If  independent information is available on S<sub>''P''</sub> =  S<sub>''E''</sub>, the additivity can be calculated for the OXPHOS state.
 |-
@@ Line 64: / Line 68: @@
 | [[S]]
 | CII
-| A succinate concentration of >10 mM may be required for saturating S<sub>''E''</sub> capacity.
+| ETS capacity with type S (CII) substrate, S<sub>''E''</sub>: ETS capacity with succinate, after blocking Complex I with rotenone. Limitation: A succinate concentration of >10 mM may be required for saturating S<sub>''E''</sub> capacity.
 |-
@@ Line 71: / Line 76: @@
 | ROX
 | ROX
-| ROX may be lower in substrate states earlier in the SUIT protocol. Therefore, this ROX measurement is frequently taken as a methodological control rather than as the basis of ROX correction of mitochondrial respiration (mt).
+| Residual oxygen consumption (ROX) due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of Complex III). ROX may be lower in substrate states earlier in the SUIT protocol. Therefore, this ROX measurement is frequently taken as a methodological control rather than as the final basis of ROX correction of mitochondrial respiration (mt).
 |}