SUIT protocol library

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Classes of protocols with coupling and substrate control

Linear

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

• CI
• CII
• CI+CII
• CIV
• ETF
• GPDH

Orthogonal

1. Orthogonal coupling -> substrate control (mono- or bidirectional)
2. Orthogonal substrate -> coupling control (mono- or bidirectional)

Diametral

1. Diametral coupling -> substrate ->> control (mono- or bidirectional)
2. Diametral substrate -> coupling ->> control (mono- or bidirectional)

CI -> CI+CII -> CII

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

• GM+D+S+F+Rot Innsbruck-Copenhagen protocol: GM+D+c+S+F+Rot

Cyt c: added after ADP; cyt c added after S; ADP conc: _ _; ROX: Ama (Boushel 2007 Diabetologia)

• PMG+D+S+F+Rot Brain protocol: PMG+D+c+S+F+Rot

ADP conc: in mouse brain homogenate a final conc. of 2 mM ADP in the OXPHOS state (CI+CII) is saturating (Sumbalova 2011 Abstract Bordeaux); ROX: Mna+Ama; Myx

CI -> CI -> CI+CII -> CII

• GM+D+P+S+F+Rot Innsbruck-Liege protocol 1: GM+D+c+P+S+F+Rot

Cytochrome c: early addition of c ensures comparability of all states in case of any c effect.

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

ETF -> CI -> CI+CII -> CII

Respiratory states: ETF_L -> ETF_P -> CI_P -> CI+II_P -> CI+II_E -> CII_E -> ROX

• OctM+D+G+S+F+Rot Innsbruck training protocol: OctM+D+G+c+S+F+Rot

Pesta 2012 Methods Mol Biol, Pesta 2011 Am J Physiol Regul Integr Comp Physiol

Respiratory states: ETF_L -> ETF_P -> CI_P -> CI+II_P -> CII_P -> CII_L -> CII_E -> ROX

• OctM+D+G+S+Rot+Omy+F Greenland-Monte Rosa protocol: OctM+D+G+S+Rot+Omy+F+c

Cyt c: added after F. 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

Experimental details

Cytochrome c

CI -> CI+CII -> CII

ETF -> CI -> CI+CII -> CII

References and Notes

• List_of_respiratory_states
• List_of_terms_and_abbreviations

• Gnaiger_2007_MitoPathways
• Gnaiger_2009_Int J Biochem Cell Biol
• Boushel 2007 Diabetologia
• Pesta 2012 Methods Mol Biol
• Pesta 2011 Am J Physiol Regul Integr Comp Physiol
• Sumbalova 2011 Abstract Bordeaux