Musiol 2018 MiPschool Tromso C4: Difference between revisions
Beno Marija (talk | contribs) (Created page with "{{Abstract |title=left|90px|Mitochondrial Physiology Society|MiPsociety |info=MitoEAGLE |authors=Musiol E |year=2018 |event=MiPschool Tromso-...") |
Beno Marija (talk | contribs) No edit summary |
||
| (4 intermediate revisions by 2 users not shown) | |||
| Line 1: | Line 1: | ||
{{Abstract | {{Abstract | ||
|title=[[Image:MiPsocietyLOGO.JPG|left|90px | |title=[[Image:MiPsocietyLOGO.JPG|left|90px|MiPsociety]] Alpha-tomatine as a novel membrane-permeabilizing agent for mitochondrial respiration measurements. | ||
|info=[[MitoEAGLE]] | |info=[[MitoEAGLE]] | ||
|authors=Musiol E | |authors=Musiol E, Weber D, Dawid C, Hofmann T, Fromme T | ||
|year=2018 | |year=2018 | ||
|event=MiPschool Tromso-Bergen 2018 | |event=MiPschool Tromso-Bergen 2018 | ||
|abstract=[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MitoEAGLE]] | |abstract=[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MitoEAGLE]] | ||
|editor=[[Beno M]], | In mitochondrial respiration measurements, digitonin is a widely used substance for the permeabilization of the plasma membrane. Digitonin is a mixture of several different saponins, naturally varying in their composition. Therefore, the effective concentration of each digitonin batch must be determined individually. We recently identified the secondary plant metabolite α-tomatine to exhibit the same permeabilizing properties as digitonin. Furthermore, it can not only permeabilize the plasma membrane but also the outer mitochondrial membrane. | ||
In this study, we used α-tomatine to selectively permeabilize the cellular and mitochondrial membrane of HEK-293 cells in respirometric measurements. The permeabilization of the outer mitochondrial membrane was combined with an excess of reduced cytochrome C to directly modulate the activity of complex IV. In standardized substrate-uncoupler-inhibitor-titration protocols, complex IV activity is usually influenced by changes in substrate availability, the substrate oxidation capacity of different enzymes and the size of the ubiquinone and cytochrome C pool. | |||
We demonstrate that our assay bypasses these limitations. Thus, the maximal activity of complex IV within the inner mitochondrial membrane can be assessed more specifically. In conclusion, the use of α-tomatine allows a more precise determination of factors responsible for changes in state 3 respiration and to increase our basic understanding of the role of complex IV in the respiratory chain. | |||
|editor=[[Beno M]], [[Plangger M]], | |||
|mipnetlab=DE Freising Klingenspor M | |||
}} | }} | ||
== Affiliations == | == Affiliations == | ||
Musiol E(1,2), Weber D(1,2), Dawid C(3), Hofmann T(3), Fromme T(1,2) | |||
::::#Chair Molecular Nutritional Medicine, Technische Univ München, Else Kröner-Fresenius Zentrum Ernährungsmedizin. - [email protected] | |||
::::#ZIEL – Inst Food Health, Technische Univ München | |||
::::#Chair Food Chemistry Molecular Sensory Science, Technische Univ München | |||
{{Labeling | |||
== | |area=Respiration, Pharmacology;toxicology | ||
|organism=Human | |||
|tissues=HEK | |||
|enzymes=Complex IV;cytochrome c oxidase | |||
|topics=Cyt c | |||
|instruments=Oxygraph-2k | |||
|event=C4, Oral | |||
}} | |||
Latest revision as of 16:54, 16 October 2018
 |
Link: MitoEAGLE
Musiol E, Weber D, Dawid C, Hofmann T, Fromme T (2018)
Event: MiPschool Tromso-Bergen 2018
In mitochondrial respiration measurements, digitonin is a widely used substance for the permeabilization of the plasma membrane. Digitonin is a mixture of several different saponins, naturally varying in their composition. Therefore, the effective concentration of each digitonin batch must be determined individually. We recently identified the secondary plant metabolite α-tomatine to exhibit the same permeabilizing properties as digitonin. Furthermore, it can not only permeabilize the plasma membrane but also the outer mitochondrial membrane.
In this study, we used α-tomatine to selectively permeabilize the cellular and mitochondrial membrane of HEK-293 cells in respirometric measurements. The permeabilization of the outer mitochondrial membrane was combined with an excess of reduced cytochrome C to directly modulate the activity of complex IV. In standardized substrate-uncoupler-inhibitor-titration protocols, complex IV activity is usually influenced by changes in substrate availability, the substrate oxidation capacity of different enzymes and the size of the ubiquinone and cytochrome C pool.
We demonstrate that our assay bypasses these limitations. Thus, the maximal activity of complex IV within the inner mitochondrial membrane can be assessed more specifically. In conclusion, the use of α-tomatine allows a more precise determination of factors responsible for changes in state 3 respiration and to increase our basic understanding of the role of complex IV in the respiratory chain.
• Bioblast editor: Beno M, Plangger M
• O2k-Network Lab: DE Freising Klingenspor M
Affiliations
Musiol E(1,2), Weber D(1,2), Dawid C(3), Hofmann T(3), Fromme T(1,2)
- Chair Molecular Nutritional Medicine, Technische Univ München, Else Kröner-Fresenius Zentrum Ernährungsmedizin. - [email protected]
- ZIEL – Inst Food Health, Technische Univ München
- Chair Food Chemistry Molecular Sensory Science, Technische Univ München
Labels: MiParea: Respiration, Pharmacology;toxicology
Organism: Human
Tissue;cell: HEK
Enzyme: Complex IV;cytochrome c oxidase Regulation: Cyt c
HRR: Oxygraph-2k
Event: C4, Oral
