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.
This page is in preparation (intern wiki: [1])
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)
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 -> LT).
- Linear substrate control: at a single coupling state (unusual).
- Monodirectional linear coupling control: LN -> PD -> EF
- Bidirectional linear coupling control: LN -> PD -> LOmy -> EF
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: CIL -> CIP -> CI&IIP -> CI&IIE -> CIIE -> 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: FAOL -> FAOP -> CI&FAOP -> CI&II&FAOP -> CI&II&FAOE -> CIIE -> 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: FAOL -> FAOP -> CI&FAOP -> CI&II&FAOP -> CIIP -> CIIL -> CIIE -> 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