SUIT protocol library

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MitoPedia

SUIT protocol library

Description

The Substrate-uncoupler-inhibitor titration (SUIT) protocol library contains a sequential list of SUIT protocols (D001, D002, ..) 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 numbers

SUIT code
SUIT-001
SUIT-002
SUIT-003
SUIT-004
SUIT-005
SUIT-006
SUIT-007
SUIT-008
SUIT-010
SUIT-011
SUIT-012
SUIT-013
SUIT-014
SUIT-015
SUIT-016
SUIT-017
SUIT-018
SUIT-019
SUIT-020
SUIT-021
SUIT-022
SUIT-024
SUIT-025
SUIT-027


List of SUIT protocols with D-numbers

SUIT codeSUIT technical name
SUIT-001 O2 ce-pce D003D003_ce1;1Dig;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
SUIT-001 O2 ce-pce D004D004_ce1;1Dig;1PM;2D;2c;3U;4G;5S;(6Oct);7Rot;8Gp;9Ama;10AsTm;11Azd
SUIT-001 O2 mt D001D001_1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
SUIT-001 O2 pfi D002D002_1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
SUIT-002 O2 ce-pce D007D007_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
SUIT-002 O2 ce-pce D007aD007a_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
SUIT-002 O2 mt D005D005_1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
SUIT-002 O2 pfi D006D006_1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd
SUIT-003 AmR ce D017D017_ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;ce7Ama
SUIT-003 AmR ce D058D058_ce1;ce1SOD;ce1HRP;ce1AmR;ce2Omy;ce3U;ce4Rot;ce5Ama
SUIT-003 AmR ce D059D059_ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D009D009_ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D012D012_ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D028D028_ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
SUIT-003 O2 ce D037D037_ce1;ce1Glc;(ce2Omy);ce3U;ce4Ama
SUIT-003 O2 ce D038D038_ce1;(ce2Omy);ce3U;ce3Glc;ce3'U;ce4Ama
SUIT-003 O2 ce D039D039_ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D050D050_ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D060D060_ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv;ce7Ama
SUIT-003 O2 ce D061D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
SUIT-003 O2 ce D062D062_ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce-pce D013D013_ce1;ce1P;ce2Omy;ce3U;ce3Glc;ce3M;ce4Rot;ce5S;1Dig;1U;1c;2Ama;3AsTm;4Azd
SUIT-003 O2 ce-pce D018D018_ce1;ce1P;ce2Omy;ce3U;ce3Glc;ce4Rot;ce5S;1Dig;1c;2Ama;3AsTm;4Azd
SUIT-003 O2 ce-pce D020D020_ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c;2Ama;3AsTm;4Azd
SUIT-004 O2 pfi D010D010_1PM;2D;2c;3U;4S;5Rot;6Ama;7AsTm;8Azd
SUIT-005 O2 pfi D011D011_1OctM;2D;3P;4S;5U;6Rot;7Ama;8AsTm;9Azd
SUIT-006 AmR mt D048D048_1PM;2D;3Omy;4U;5Ama
SUIT-006 Fluo mt D034D034_1PM;2D;3Omy;4U;5Ama
SUIT-006 MgG mt D055D055_1PM;2D;3Cat;4U;5Ama
SUIT-006 O2 ce-pce D029D029_ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama
SUIT-006 O2 mt D022D022_1SRot;2D;2c;(3Omy);4U;5Ama
SUIT-006 O2 mt D047D047_1PM;2D;2c;3Omy;4U;5Ama
SUIT-007 O2 ce-pce D030D030_ce1;1Dig;1G;2D;2c;3M;4U;5Ama
SUIT-008 O2 ce-pce D025D025_ce1;1Dig;1PM;2D;2c;3G;4S;5U;6Rot;7Ama;8AsTm;9Azd
SUIT-008 O2 mt D026D026_1PM;2D;2c;3G;4S;4D;5U;6Rot;7Ama;8AsTm;9Azd
SUIT-008 O2 pfi D014D014_1PM;2D;2c;3G;4S;4D;5U;6Rot;7Ama;8AsTm;9Azd
SUIT-009 AmR ce-pce D019D019_ce1;1Dig;1S;2D;3P;4Rot;5Ama
SUIT-009 AmR mt D021D021_1S;2D;3P;4Rot;5Ama
SUIT-009 O2 ce-pce D016D016_ce1;1Dig;1S;2D;3P;4Rot;5Ama
SUIT-009 O2 mt D015D015_1S;2D;3P;4Rot;5Ama
SUIT-010 O2 ce-pce D008D008_ce1;ce2Rot;ce3S;ce4D;1Dig;1c
SUIT-011 O2 pfi D024D024_1GM;2D;2c;3S;4U;5Rot;6Ama
SUIT-012 O2 ce-pce D052D052_ce1;1Dig;1PM;2D;2c;3G;4U;5Ama
SUIT-012 O2 mt D027D027_1PM;2D;2c;3G;4U;5Ama
SUIT-013 AmR ce D023D023_ce1
SUIT-014 O2 pfi D042D042_1GM;2D;3P;4S;5U;6Rot;7Ama
SUIT-015 O2 pti D043D043_1OctM;2D;3G;4P;5S;6U;7Rot;8Ama
SUIT-016 O2 pfi D044D044_1OctM;2D;3G;4S;5Rot;6Omy;7U;8Ama
SUIT-017 O2 mt D046D046_1OctM;2D;3G;3c;4S;5U;6Rot;7Ama
SUIT-017 O2 pfi D049D049_1OctM;2D;2c;3G;4S;5U;6Rot;7Ama
SUIT-018 AmR mt D031D031_1GMS;2D;3Ama
SUIT-018 AmR mt D040D040_1GMS;2D;3Ama
SUIT-018 AmR mt D041D041_1GMS;2D;3Ama
SUIT-018 O2 mt D054D054_1GMS;2D;3Ama
SUIT-019 O2 pfi D045D045_1PalM;2D;2c;3Oct;4P;5G;6U;7S;8Rot;9Ama
SUIT-020 Fluo mt D033D033_1PM;2D;3G;4S;5Rot;6Omy;7U;8Ama
SUIT-020 O2 mt D032D032_1PM;2D;2c;3G;4S;5Rot;6Omy;7U;8Ama
SUIT-021 Fluo mt D036D036_1GM;2D;3S;4Rot;5Omy;6U;7Ama
SUIT-021 O2 mt D035D035_1GM;2D;2c;3S;4Rot;5Omy;6U;7Ama
SUIT-022 O2 ce D051D051_ce1;ce2KCN;ce3SHAM
SUIT-023 O2 ce D053D053_ce1;ce2SHAM;ce3KCN
SUIT-024 O2 ce-pce D056D056_ce1;1Dig;1PM;2T;2D;3Omy;4Ama
SUIT-025 O2 mt D057D057_1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot;9Ama
SUIT-026 AmR mt D064D064_1S;2Rot;3D;4Ama
SUIT-026 O2 mt D063D063_1S;2Rot;3D;3c;4Ama
SUIT-027 O2 ce-pce D065D065_ce1;1Dig;1M;2D;3M;4P;5G

Strategy

  1. Define substrates and inhibitors for each pathway state.
  2. 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.

Coupling states: LEAK OXPHOS ET-capacity


Define residual oxygen consumption

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

ROX


Categories of SUIT protocols

» Categories of SUIT protocols
» Talk:SUIT protocol library

SUIT protocol patterns

» SUIT protocol pattern


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.
  • 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</sub> -> ROX


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