SUIT-004 O2 pfi D010

high-resolution terminology - matching measurements at high-resolution

SUIT-004 O2 pfi D010

Description

Abbreviation: NS(PM)

Reference: A: RP1m (RP1 modified) is a short SUIT protocol linked to 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp- - SUIT RP1 (human skeletal muscle)

MitoPedia concepts: SUIT protocol, SUIT A 

SUIT-category: NS(PM)

SUIT protocol pattern: orthogonal

• Linked to 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp- - SUIT RP1, specifically for human skeletal muscle mitochondria.
• Harmonized with 1OctM;2D;3P;4S;5U;6Rot-.

1PM;2D;2c;3U;4S;5Rot-

300px 1PM;2D;2c;3U;4S;5Rot-

Specific cases

• imt, ROX: NS(PM)01_imt,1PM,2D(c),3U,4S,5Rot,6Ama
• pfi, high O₂, no ROX: NS(PM)01_pfiO2,1PM,2D(c),3U,4S,5Rot

Link to RP1

The 1PM;2D;3U;4S;5Rot- protocol is linked to 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp- for healthy human skeletal muscle mitochondria on the basis of the following rationale.

1. The first coupling control steps (1-4) are identical.
2. RP1-5G is skipped, since PM and PGM yield nearly identical OXPHOS capacity. However, PM<PGM is a potential diagnostic for a defect in the PM-linked pathway upstream of CI.
3. RP1-7Oct is skipped, since octanoylcarnitine exerts practically no stimulatory effect on OXPHOS capacity in state PMS or PGMS, but in conjunction with prolonged exposure may even yield a slight decline of respiration.
4. RP1-9Gp is skipped, since glycerolphosphate exerts only a minor stimulatory effect on OXPHOS capacity in state SRot.
5. Omission of the three steps described above shortens the SUIT protocol, thus reducing the risk of diminishing respiratory capacity over time, reducing the number of necessary reoxygenations, and providing more time for detailed evaluation of steady-states. The loss of diagnostic information may be minor relative to the benefits of simplification of the protocol, particularly in studies of exercise in healthy humans.
6. RP1-10Ama may be skipped, since correction of the initial states by ROX measured in RP1-10 Ama may represent an over-correction, and inhibition takes a very long time in several cases (Pesta 2012 Methods Mol Biol).
7. RP1-11Tm and RP1-12Azd are skipped, if a prolonged protocol is not practical due to limination of time.

SUIT O2 DLP for RP1m (DatLab 7.1)

DL-Protocols (DLP) for substrate-uncoupler-inhibitor titrations (see MitoPedia: SUIT) guide through a sequence of coupling and pathway control states. A specific SUIT protocol can be assigned to O2k-Chamber A or B or both. DL-Protocols for different SUIT protocols can be found in the library of protocols MitoPedia: SUIT. Instrumental DL-protocols are displayed in the DL-Protocol library.

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DL-Protocol	Description	Download - Protocol .DLP	Download - Analysis .XLSX
RP1m	Modified SUIT reference protocol RP1 for permeabilized fibers