SUIT protocol library: Difference between revisions
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== List of SUIT protocol D-numbers == | == List of SUIT protocol D-numbers == | ||
{{#ask:[[MitoPedia concept::SUIT A]] | {{#ask:[[MitoPedia concept::SUIT A]] [[MitoPedia concept::SUIT D]] | ||
| ?Has SUIT number=SUIT technical name | | ?Has SUIT number=SUIT technical name | ||
| mainlabel=SUIT code | | mainlabel=SUIT code |
Revision as of 12:13, 11 January 2019
Description
The Substrate-uncoupler-inhibitor titration (SUIT) protocol library contains a sequential list of SUIT protocols (D01, D02, ..) with links to the specific SUIT pages. Classes of SUIT protocols are explained with coupling and substrate control defined for mitochondrial preparations.
Abbreviation: SUITs
Reference: MitoPedia: SUIT
List of SUIT protocol D-numbers
Strategy
- 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)
Define residual oxygen consumption
- ROX: Rot&Ama, Rot&Mna&Ama, Myx, Cn, Az, ..
Categories of SUIT protocols
SUIT protocol patterns
Examples of SUIT protocols
N; NS; S diametral
- Old nomenclature: CI + CI&II + CII diametral
- Respiratory states: NL -> NP -> NSP -> NSE -> SE -> 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 (NSP) 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.
- Old nomenclature: CI + CI + CI&II + CII diamteral
- 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: FL -> FP -> FNP -> FNSP -> FNSE -> 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: FL -> FP -> FNP -> FNSP -> SP -> SL -> SE -> 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
MitoPedia concepts:
MiP concept,
SUIT concept