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

Coupling states:

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 -> L_T).
Linear substrate control: at a single coupling state (unusual).
Monodirectional linear coupling control: L_N -> P_D -> E_F
Bidirectional linear coupling control: L_N -> P_D -> L_Omy -> E_F