SUIT protocol library

high-resolution terminology - matching measurements at high-resolution

SUIT protocol library

Description

The Substrate-uncoupler-inhibitor titration (SUIT) protocol library contains classes of SUIT protocols with coupling and substrate control defined for mitochondrial preparations.

Abbreviation: SUITs

MitoPedia concepts: "MitoFit Quality Control System" is not in the list (MiP concept, Respiratory state, Respiratory control ratio, SUIT concept, SUIT protocol, SUIT A, SUIT B, SUIT C, SUIT state, Recommended, ...) of allowed values for the "MitoPedia concept" property. MitoFit Quality Control System"MitoFit Quality Control System" is not in the list (Enzyme, Medium, Inhibitor, Substrate and metabolite, Uncoupler, Sample preparation, Permeabilization agent, EAGLE, MitoGlobal Organizations, MitoGlobal Centres, ...) of allowed values for the "MitoPedia topic" property.

Template:MitoPedia SUIT

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Strategy

Select a pathway control state.
Select a control pattern.
Define substrates for each pathway state.
Define coupling states.

Definition of states

Experimental conditions

Temp
O2
Medium
Oxygen level
ADP kinetics
Substrate kinetics

O2k configuration

Amp: Amplex Red, TMRM, Ca Green, ...
Pot
Spectrophotometry

Define substrates

Substrate control states are defined for each pathway control state (CI-linked, CII-linked, CI&II-linked) is defined by a specific substrate or substrate combination. These substrates may be added in a single step, or in multiple steps for analyis of the effect of different substrate combinations within a pathway control state.

CI: PM, GM, PGM, M, ..
CII: S(Rot), S, SM, ..
CI&II: PS, GS, PMS, GMS, PGMS, ..
CIV: AsTM
FAO: Oct, Pal, ...
CGpDH:
c

Define coupling states

The coupling control states require a complementary definition of an ETS-competent substrate state.

L, P, E (Uncouplers, U: FCCP, CCCP, DNP)

Coupling states:

Define residual oxygen consumption

ROX: Rot&Ama, Rot&Mna&Ama, Myx, Cn, Az, ..

Pathway control states

Single pathway control states

CI
CII
CI&II
CIV
FAO
CGpDH

Multiple pathway control states

A. CI-CII-CIV

CI + CII
CI + CII + CIV
CI + CI&II + CII
CI + CI&II + CII + CIV

B. FAO-CI-CII-CGpDH-CIV

FAO + CI&FAO + CII + CGpDH
FAO + CI&FAO + CII + CGpDH + CIV
FAO + CI&FAO + CI&II&FAO + CII + CGpDH
FAO + CI&FAO + CI&II&FAO + CII + CGpDH + CIV

C. FAO-CI-CII-CIV

FAO + CI&FAO + CII
FAO + CI&FAO + CII + CIV
FAO + CI&FAO + CI&II&FAO + CII
FAO + CI&FAO + CI&II&FAO + CII + CIV

D. CGpDH-CI-CII-CIV

CGpDH + CI&GpDH
CGpDH + CI&GpDH + CII&GpDH
CGpDH + CI&GpDH + CII&GpDH + CIV
CGpDH + CI&GpDH + CI&II&GpDH + CII&GpDH
CGpDH + CI&GpDH + CI&II&GpDH + CII&GpDH + CIV
CI + CI&GpDH + ...
CI + CI&II + CI&II&GpDH + ...
CI&II + CI&II&GpDH + ...

E. FAO-CGpDH-CI-CII-CIV

FAO + FAO&GpDH + CI&FAO&GpDH + ...
CGpDH + FAO&GpDH + CI&FAO&GpDH + ...
FAO + FAO&GpDH + CI&II&FAO + CI&II&FAO&GpDH + ...
CGpDH + FAO&GpDH + CI&FAO&GpDH + ...
CI + CI&II + CI&II&GpDH + CI&II&FAO&GpDH + ...
...

Control patterns

Linear

Linear coupling control: at a single substrate state (classical State 3 -> State 4, or P -> L_T).
Linear substrate control: at a single coupling state (unusual).
Monodirectional linear coupling control: L_N -> P_D -> E_F
Bidirectional linear coupling control: L_N -> P_D -> L_Omy -> E_F

Orthogonal

Orthogonal coupling -> substrate control (mono- or bidirectional)
Orthogonal substrate -> coupling control (mono- or bidirectional)

Diametral

Diametral coupling -> substrate ->> control (mono- or bidirectional)
Diametral substrate -> coupling ->> control (mono- or bidirectional)

SUIT library

CI + CI&II + CII diametral

Respiratory states: CI_L -> CI_P -> CI&II_P -> CI&II_E -> CII_E -> ROX

GM+D+S+U+Rot Innsbruck-Copenhagen protocol: GM+D+c+S+U+Rot ::Cyt c: added after ADP; cyt c added after S; ADP conc: _ _; ROX: Ama (Boushel 2007 Diabetologia)

PMG+D+S+U+Rot Brain protocol: PMG+D+c+S+U+Rot::ADP conc: in mouse brain homogenate a final conc. of 2 mM ADP in the OXPHOS state (CI&II) is saturating (Sumbalova 2011 Abstract Bordeaux); ROX: Mna&Ama; Myx

CI + CI + CI&II + CII diamteral

GM+D+P+S+U+Rot Innsbruck-Liege protocol 1: GM+D+c+P+S+U+Rot::Cytochrome c: early addition of c ensures comparability of all states in case of any c effect.

PM+D+G+S+U+Rot Innsbruck-Liege protocol 2: PM+D+c+G+S+U+Rot

FAO + CI + CI&II + CII diametral

Respiratory states: FAO_L -> FAO_P -> CI&FAO_P -> CI&II&FAO_P -> CI&II&FAO_E -> CII_E -> ROX

OctM+D+G+S+U+Rot Innsbruck training protocol: OctM+D+G+c+S+U+Rot::Pesta 2012 Methods Mol Biol, Pesta 2011 Am J Physiol Regul Integr Comp Physiol

Respiratory states: FAO_L -> FAO_P -> CI&FAO_P -> CI&II&FAO_P -> CII_P -> CII_L -> CII_E -> ROX

OctM+D+G+S+Rot+Omy+U Greenland-Monte Rosa protocol: OctM+D+G+S+Rot+Omy+U+c::Cyt c: added after U. The absence of a significant c-effect shows the integrity of the outer mitochondrial membrane after fibre preparation and also shows the preservation of integrity over a 90-100 min incubation in the O2k (MiR05); Oxygen: oxygen limitation of flux was preventd by maintaining oxygen levels in the respirometer above air saturation (Pesta 2012 Methods Mol Biol); ROX: Mna+Ama

References and notes