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

Reference: MitoPedia: SUIT

MitoPedia concepts: MiP concept, SUIT concept

This page is in preparation (intern wiki: [1])

Strategy

Select a pathway control state.
Select a control pattern.
Define substrates and inhibitors 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 pathway states

Pathway control states are defined for many substrate-inhibitor combinations in SUIT protocols, particularly: (N-pathway control state, S-pathway control state, NS-pathway control state). Titrations may performed in a single step, or in multiple steps for sequential analyis of different pathway control states.

N: PM, GM, PGM, M, ..
S: S(Rot), S, SM, ..
NS: PS, GS, PMS, GMS, PGMS, ..
CIV: AsTM
F: Oct, Pal, ...
Gp

Define coupling states

The coupling control states require a complementary definition of an ET-pathway competent state.

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

Coupling states:

Define residual oxygen consumption

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

Categories of SUIT protocols

» Categories of SUIT protocols

» Talk:SUIT protocol library

SUIT protocol patterns

» SUIT protocol pattern

SUIT library

N; NS; S diametral

Old nomenclature: CI + CI&II + CII diametral

Respiratory states: N_L -> N_P -> NS_P -> NS_E -> S_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: PGM;D;c;S;U;Rot::ADP conc: in mouse brain homogenate a final conc. of 2 mM ADP in the NS-OXPHOS state (NS_P) is saturating (Sumbalova 2011 Abstract Bordeaux); ROX: Mna&Ama; Myx

N; N; NS; S diamteral

Old nomenclature: 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

F; FN; FNS; S diametral

Old nomenclature: FAO + CI + CI&II + CII diametral

Respiratory states: F_L -> F_P -> FN_P -> FNS_P -> FNS_E -> SE -> 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: F_L -> F_P -> FN_P -> FNS_P -> S_P -> S_L -> S_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