Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

Difference between revisions of "SUIT-001 O2 ce-pce D004"

From Bioblast
Line 1: Line 1:
{{MitoPedia
{{MitoPedia
|abbr=NSGp(PGM)
|description=[[File:ce1;1Dig;1PM;2D;2c;3U;4G;5S;7Rot;8Gp;9Ama;10AsTm;11Azd.png|400px|SUIT-RP1 for PBMCs and PLTs]]
|info='''A''' Reference protocol RP1; pce: specific for permeabilized PBMC and PTL cells - '''[[SUIT-001]]'''
|application=O2
|application=O2
|description=[[File:ce1;1Dig;1PM;2D;2c;3U;4G;5S;7Rot;8Gp-.png|400px|SUIT-RP1 for PBMCs and PLTs]]
|abbr=NSGp(PGM)
|SUIT number=D004_ce1;1Dig;1PM;2D;2c;3U;4G;5S;7Rot;8Gp;9Ama;10AsTm;11Azd  
|SUIT number=D004_ce1;1Dig;1PM;2D;2c;3U;4G;5S;7Rot;8Gp;9Ama;10AsTm;11Azd  
|info='''A''' Reference protocol RP1 specific for PBMC and PTL cells''' - [[SUIT-001]]
}}
}}
::: '''[[MitoPedia: SUIT]]''' - '''[[SUIT reference protocol]] RP1 for PBMCs and PLTs'''
::: '''[[MitoPedia: SUIT]]''' - '''[[SUIT reference protocol]] RP1 for PBMCs and PLTs'''
Line 10: Line 10:
::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;2c;3U;4G;5S;7Rot;8Gp-
::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;2c;3U;4G;5S;7Rot;8Gp-


::::* Reference protocol 1 specifically for PBMCs and PLTs.  
The SUIT-001 O2 pce D004 protocol is the reference protocol 1 (RP1) specifically for PBMCs and PLTs. The main difference between the RP1 for PBMCs and PLTs and RP1 for other mitochondrial preparations [[SUIT-001| see SUIT-001]] is the absence of Oct in the case of blood cells.  
::::* The main difference between the RP1 for PBMCs and PLTs and RP1 for other mtprep [[1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp-|SUIT RP1]] is the absence of Oct in the case of blood cells.  
 
The SUIT-001 O2 pce D004 protocol in combination with [[SUIT-002_O2_pce_D007a]] is specially designed to provide a common reference for comparison of respiratory control of [[PBMC|PBMC]] and [[Platelet|platelets]] cells in a wide variety of species. SUIT-001 O2 pce D004 gives information of of [[ROUTINE]] respiration (intact cells) and after the plasma membrane permeabilization (permeabilized cells) of the linear coupling control ([[LEAK-respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH linked-substrates ([[PM pathway control state|PM]]). Moreover, the pathway control in ET state ([[NADH_Electron_transfer-pathway_state|N]], [[NS-pathway control state| NS]], [[Succinate pathway control state| S]] and [[SGp-pathway control state| SGp]] pathways) can be evaluated by using this SUIT protocol. SUIT-001 O2 pce D004 can be extended with the CIV assay module.
The SUIT-001 O2 pce D004 protocol in combination with [[SUIT-002_O2_pce_D007a]] is specially designed to provide a common reference for comparison of respiratory control of [[PBMC|PBMC]] and [[Platelet|platelets]] cells in a wide variety of species. SUIT-001 O2 pce D004 gives information of of [[ROUTINE]] respiration (intact cells) and after the plasma membrane permeabilization (permeabilized cells) of the linear coupling control ([[LEAK-respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH linked-substrates ([[PM pathway control state|PM]]). Moreover, the pathway control in ET state ([[NADH_Electron_transfer-pathway_state|N]], [[NS-pathway control state| NS]], [[Succinate pathway control state| S]] and [[SGp-pathway control state| SGp]] pathways) can be evaluated by using this SUIT protocol. SUIT-001 O2 pce D004 can be extended with the CIV assay module.



Revision as of 12:45, 6 March 2019


high-resolution terminology - matching measurements at high-resolution


SUIT-001 O2 ce-pce D004

Description

SUIT-RP1 for PBMCs and PLTs

Abbreviation: NSGp(PGM)

Reference: A Reference protocol RP1; pce: specific for permeabilized PBMC and PTL cells - SUIT-001

SUIT number: D004_ce1;1Dig;1PM;2D;2c;3U;4G;5S;7Rot;8Gp;9Ama;10AsTm;11Azd

O2k-Application: O2

MitoPedia: SUIT - SUIT reference protocol RP1 for PBMCs and PLTs
SUIT-category: NSGp(PGM)
SUIT protocol pattern: orthogonal 1PM;2D;2c;3U;4G;5S;7Rot;8Gp-

The SUIT-001 O2 pce D004 protocol is the reference protocol 1 (RP1) specifically for PBMCs and PLTs. The main difference between the RP1 for PBMCs and PLTs and RP1 for other mitochondrial preparations see SUIT-001 is the absence of Oct in the case of blood cells. The SUIT-001 O2 pce D004 protocol in combination with SUIT-002_O2_pce_D007a is specially designed to provide a common reference for comparison of respiratory control of PBMC and platelets cells in a wide variety of species. SUIT-001 O2 pce D004 gives information of of ROUTINE respiration (intact cells) and after the plasma membrane permeabilization (permeabilized cells) of the linear coupling control (L- P- E) with NADH linked-substrates (PM). Moreover, the pathway control in ET state (N, NS, S and SGp pathways) can be evaluated by using this SUIT protocol. SUIT-001 O2 pce D004 can be extended with the CIV assay module.


Communicated by Doerrier C and Gnaiger E (last update 2019-03-06)
MitoPedia: SUIT

Steps and respiratory states

SUIT-001 for PBMCs and PLTs

Step State Pathway Q-junction Comment - Events (E) and Marks (M)
ce1 ROUTINE ce1
  • ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
1Dig REN ce1;1Dig
  • Optimum effective digitonin concentration for complete plasma membrane permeabilization.
Step State Pathway Q-junction Comment - Events (E) and Marks (M)
1PM PML(n) N CI PML or PM_L: Pyruvate & Malate, N-LEAK respiration, NL
2D PMP N CI 1PM;2D
2c PMcP N CI 1PM;2D;2c
3U PME N CI 1PM;2D;3U
4G PGME N CI 1PM;2D;3U;4G
5S PGMSE NS CI&II 1PM;2D;3U;4G;5S
(6Oct) 1PM;2D;3U;4G;5S;(6Oct)
  • Oct is skipped in PBMCs and PTLs
7Rot SE S CII 1PM;2D;3U;4G;5S;(6Oct);7Rot
8Gp SGpE SGp CII&GpDH 1PM;2D;3U;4G;5S;(6Oct);7Rot;8Gp
  • Respiratory stimulation by action of succinate and glycerophosphate, Gp, with convergent electron flow in the SGp-pathway (CII&GpDH-linked pathway to the Q-junction).
  • Noncoupled electron transfer state, ET state, with ET capacity E.
9Ama ROX
  • Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).
Step Respiratory state Pathway control ET-Complex Comment
## AsTm AsTmE CIV CIV
## Azd CHB


Questions.jpg


Click to expand or collaps

Strengths and limitations

  • SUIT-001 in combination with SUIT-002 provides a common reference for comparison of respiratory control in a large variety of species, tissues and cell types. Both SUIT protocols provide a mitochondrial mapping which allows:
1. to obtain reference values.
2. to evaluate mitochondrial physiological diversity, generating a mt-database on comparative mitochondrial physiology.
3. to screen specific defects.
  • SUIT-001 and SUIT-002 are used in the MitoFit Proficiency test for inter-individual and inter-laboratory reproducibility quality control.
  • A succinate concentration of >10 mM may be required for saturating SE capacity.
+ Linear coupling control (L-P-E) with N substrates (PM).
+ Pathway control in ET state (N, NS, FNS, S and SGp pathways).
+ Harmonization with SUIT-002 allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated measurements.
+ In blood cells, SUIT-001 and SUIT-002 harmonize in: ROUTINE (ce1), SGpE (8Gp in SUIT-001 and 10Rot in SUIT-002) and Complex IV.
+ Harmonization with many SUIT protocols (up to step 5S).
+ This protocol can be extended with the Complex IV module.
- The substrate combination for maximum ET capacity (FNSGP) is not obtained in SUIT-001 in favour of measuring SE.
- Oct is not added in ET state.
- Lengthy duration of the experiment.

Compare SUIT protocols

  • SUIT-001 O2 pce D004 (SUIT-001 for permeabilized PBMCs and PLTs) differs from SUIT-001 O2 pce D003 (general SUIT-001 for permeabilized cells) in the absence and presence of octanoylcarnitine, respectively.
  • SUIT-004: Short version of RP1; without G, F and Gp.
  • SUIT-006: Very short initial coupling control module.

References

MitoPedia concepts: SUIT protocol, SUIT A, Find 


MitoPedia methods: Respirometry