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Difference between revisions of "Coupling-control protocol"

From Bioblast
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{{MitoPedia
{{MitoPedia
|abbr=PCP
|abbr=PCP
|description=A '''phosphorylation control protocol''' induces different [[coupling control state]]s at constant substrate supply. In [[intact cell]]s, the PCP can be applied by using membrane-permeable inhibitors of the [[phosphorylation system]] (e.g. [[oligomycin]]) and [[uncoupler]]s (e.g. [[CCP]]). Coupling control states in intact cells include ''R'', ''L'', ''E''; [[LEAK]], [[ROUTINE]], and [[ETS]]. Coupling control states in isolated mitochondria, permeabilized cells or homogenates include ''L'', ''P'', ''E''; LEAK, [[OXPHOS]], and ETS.
|description=A '''phosphorylation control protocol''' induces different [[coupling control state]]s at constant substrate supply. In [[intact cell]]s, the PCP can be applied by using membrane-permeable inhibitors of the [[phosphorylation system]] (e.g. [[oligomycin]]) and [[uncoupler]]s (e.g. [[CCCP]]). Coupling control states in intact cells include ''R'', ''L'', ''E''; [[LEAK]], [[ROUTINE]], and [[ETS]]. Coupling control states in isolated mitochondria, permeabilized cells or homogenates include ''L'', ''P'', ''E''; LEAK, [[OXPHOS]], and ETS.
|info=[[Pesta 2012 Methods Mol. Biol.]]
|info=[[Gnaiger 2008 POS]]
|type=Respiration
}}
}}
{{MitoPedia methods
{{MitoPedia methods
|mitopedia method=Respirometry
|mitopedia method=Respirometry
|type=Respiration
}}
}}
{{MitoPedia topics
{{MitoPedia topics
Line 13: Line 11:
|type=Respiration
|type=Respiration
}}
}}
== From PCP to CCP ==
In functional OXPHOS analysis, the control of oxidative phosphorylation by coupling is of primary importance, as studied by application of protocols, in which the [[phosphorylation system]] is either inhibited (lack of ADP; inhibition by oligomycin), activated (saturating ADP; activation by phyiological control in intact cells) or eliminated (uncoupling). It seems thus appropriate to use the term phosphorylation control protocol, PCP <ref> Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley:327-52. »[[Gnaiger_2008_POS |Bioblast Access]]« </ref>. In this context it is taken for granted that we do not refer to metabolic control by phosphorylation of enzymes, which is an important mechanism to change specific enzyme activity. To avoid any such confusion, it is suggested to replace the term 'phosphorylation control protocol' by '''coupling control protocol, CCP'''
<ref> Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. OROBOROS MiPNet Publications, Innsbruck:80 pp. »[[Gnaiger 2014 MitoPathways |Open Access]]« </ref>
[[Image:R.jpg|link=ROUTINE respiration|ROUTINE]] - [[Image:L.jpg|link=LEAK respiration|LEAK]] - [[Image:E.jpg|link=ETS capacity|ETS]] - [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]]: this is the coupling control protocol with intact cells (ROUTINE - LEAK<sub>Omy</sub> - ETS - ROX).
[[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] - [[Image:L.jpg|link=LEAK respiration|LEAK]] - [[Image:E.jpg|link=ETS capacity|ETS]] - [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]]: this is the comparable coupling control protocol with [[mitochondrial preparations]] (OXPHOS - LEAK<sub>Omy</sub> - ETS - ROX).
[[Image:L.jpg|link=LEAK respiration|LEAK]] - [[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] - [[Image:L.jpg|link=LEAK respiration|LEAK]] - [[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] - [[Image:E.jpg|link=ETS capacity|ETS]] - [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]]: In mitochondrial preparations, various variations are possible for the coupling control protocol, for example in isolated mitochondria
<ref> Hand SC, Gnaiger E (2014) Flux control ratios in isolatd mitochondria. OXPHOS capacity and respiratory control in isolated mitochondria. Mitochondr Physiol Network 12.11(06):1-5. »[MiPNet12.11 MitoRespiration |Open Access]]« </ref>: (LEAK<sub>N</sub> - OXPHOS - LEAK<sub>T</sub> - OXPHOS - ETS - ROX).<ref> </ref>
<references/>
== Biochemical coupling efficiency: from 0 to <1 ==
== Biochemical coupling efficiency: from 0 to <1 ==
* ''More details:'' [[ETS coupling efficiency]]
* ''More details:'' [[ETS coupling efficiency]]

Revision as of 11:23, 11 January 2015


high-resolution terminology - matching measurements at high-resolution


Coupling-control protocol

Description

A phosphorylation control protocol induces different coupling control states at constant substrate supply. In intact cells, the PCP can be applied by using membrane-permeable inhibitors of the phosphorylation system (e.g. oligomycin) and uncouplers (e.g. CCCP). Coupling control states in intact cells include R, L, E; LEAK, ROUTINE, and ETS. Coupling control states in isolated mitochondria, permeabilized cells or homogenates include L, P, E; LEAK, OXPHOS, and ETS.

Abbreviation: PCP

Reference: Gnaiger 2008 POS


MitoPedia methods: Respirometry 


MitoPedia topics: "Respiratory state" is not in the list (Enzyme, Medium, Inhibitor, Substrate and metabolite, Uncoupler, Sample preparation, Permeabilization agent, EAGLE, MitoGlobal Organizations, MitoGlobal Centres, ...) of allowed values for the "MitoPedia topic" property. Respiratory state"Respiratory state" is not in the list (Enzyme, Medium, Inhibitor, Substrate and metabolite, Uncoupler, Sample preparation, Permeabilization agent, EAGLE, MitoGlobal Organizations, MitoGlobal Centres, ...) of allowed values for the "MitoPedia topic" property. 

From PCP to CCP

In functional OXPHOS analysis, the control of oxidative phosphorylation by coupling is of primary importance, as studied by application of protocols, in which the phosphorylation system is either inhibited (lack of ADP; inhibition by oligomycin), activated (saturating ADP; activation by phyiological control in intact cells) or eliminated (uncoupling). It seems thus appropriate to use the term phosphorylation control protocol, PCP [1]. In this context it is taken for granted that we do not refer to metabolic control by phosphorylation of enzymes, which is an important mechanism to change specific enzyme activity. To avoid any such confusion, it is suggested to replace the term 'phosphorylation control protocol' by coupling control protocol, CCP [2]


ROUTINE - LEAK - ETS - ROX: this is the coupling control protocol with intact cells (ROUTINE - LEAKOmy - ETS - ROX).

OXPHOS - LEAK - ETS - ROX: this is the comparable coupling control protocol with mitochondrial preparations (OXPHOS - LEAKOmy - ETS - ROX).

LEAK - OXPHOS - LEAK - OXPHOS - ETS - ROX: In mitochondrial preparations, various variations are possible for the coupling control protocol, for example in isolated mitochondria [3]: (LEAKN - OXPHOS - LEAKT - OXPHOS - ETS - ROX).Cite error: Invalid <ref> tag; refs with no name must have content

  1. Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley:327-52. »Bioblast Access« 
  2. Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. OROBOROS MiPNet Publications, Innsbruck:80 pp. »Open Access« 
  3. Hand SC, Gnaiger E (2014) Flux control ratios in isolatd mitochondria. OXPHOS capacity and respiratory control in isolated mitochondria. Mitochondr Physiol Network 12.11(06):1-5. »[MiPNet12.11 MitoRespiration |Open Access]]« 

Biochemical coupling efficiency: from 0 to <1