SUIT protocol library
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Substrate-uncoupler-inhibitor titration (SUIT) protocols
Classes of SUIT protocols with coupling and substrate control defined for mitochondrial preparations
Strategy
- Select a pathway control state.
- Select a control pattern.
- Define substrates for each pathway state.
- Define coupling states.
Definition of states
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
Define residual oxygen consumption
- ROX: Rot&Ama, Rot&Mna&Ama, Cn, Az, ..
Pathway control states
Single pathway control states
- CI
- CII
- CI&CII
- CIV
- FAO
- CGPDH
Multiple pathway control states
A. CI-CII-CIV
- CI + CII
- CI + CII + CIV
- CI + CI&CII + CII
- CI + CI&CII + CII + CIV
B. FAO-CI-CII-CIV
- FAO + CI&FAO + CII
- FAO + CI&FAO + CII + CIV
- FAO + CI&FAO + CI&CII&FAO + CII
- FAO + CI&FAO + CI&CII&FAO + CII + CIV
C. CGPDH-CI-CII-CIV
- CGPDH + CI&CGPDH
- CGPDH + CI&CGPDH + CII&CGPDH
- CGPDH + CI&CGPDH + CII&CGPDH + CIV
- CGPDH + CI&CGPDH + CI&CII&CGPDH + CII&CGPDH
- CGPDH + CI&CGPDH + CI&CII&CGPDH + CII&CGPDH + CIV
- CI + CI&CGPDH + ...
- CI + CI&CII + CI&CII&CGPDH + ...
- CI&CII + CI&CII&CGPDH + ...
D. FAO-CGPDH-CI-CII-CIV
- FAO + FAO&CGPDH + CI&FAO&CGPDH + ...
- CGPDH + FAO&CGPDH + CI&FAO&CGPDH + ...
- CI + CI&FAO + CI&FAO&CGPDH + ...
- CI + CI&CGPDH + CI&FAO&CGPDH + ...
- ...
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)