Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization.

2.50
Hdl Handle:
http://hdl.handle.net/10033/621128
Title:
Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization.
Authors:
Nonnenmacher, Yannic; Palorini, Roberta; d'Herouël, Aymeric Fouquier; Krämer, Lisa; Neumann-Schaal, Meina; Chiaradonna, Ferdinando; Skupin, Alexander; Wegner, Andre; Hiller, Karsten
Abstract:
To date, it is well-established that mitochondrial dysfunction does not only play a vital role in cancer but also in other pathological conditions such as neurodegenerative diseases and inflammation. An important tool for the analysis of cellular metabolism is the application of stable isotope labeled substrates, which allow for the tracing of atoms throughout metabolic networks. While such analyses yield very detailed information about intracellular fluxes, the determination of compartment specific fluxes is far more challenging. Most approaches for the deconvolution of compartmented metabolism use computational models whereas experimental methods are rare. Here, we developed an experimental setup based on selective permeabilization of the cytosolic membrane that allows for the administration of stable isotope labeled substrates directly to mitochondria. We demonstrate how this approach can be used to infer metabolic changes in mitochondria induced by either chemical or genetic perturbations and give an outlook on its potential applications.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization. 2017, 43 (Pt B):147-155 Metab. Eng.
Journal:
Metabolic engineering
Issue Date:
Sep-2017
URI:
http://hdl.handle.net/10033/621128
DOI:
10.1016/j.ymben.2016.12.005
PubMed ID:
27988388
Type:
Article
Language:
en
ISSN:
1096-7184
Appears in Collections:
publications of the scientific administration (GFW)

Full metadata record

DC FieldValue Language
dc.contributor.authorNonnenmacher, Yannicen
dc.contributor.authorPalorini, Robertaen
dc.contributor.authord'Herouël, Aymeric Fouquieren
dc.contributor.authorKrämer, Lisaen
dc.contributor.authorNeumann-Schaal, Meinaen
dc.contributor.authorChiaradonna, Ferdinandoen
dc.contributor.authorSkupin, Alexanderen
dc.contributor.authorWegner, Andreen
dc.contributor.authorHiller, Karstenen
dc.date.accessioned2017-10-06T09:18:42Z-
dc.date.available2017-10-06T09:18:42Z-
dc.date.issued2017-09-
dc.identifier.citationAnalysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization. 2017, 43 (Pt B):147-155 Metab. Eng.en
dc.identifier.issn1096-7184-
dc.identifier.pmid27988388-
dc.identifier.doi10.1016/j.ymben.2016.12.005-
dc.identifier.urihttp://hdl.handle.net/10033/621128-
dc.description.abstractTo date, it is well-established that mitochondrial dysfunction does not only play a vital role in cancer but also in other pathological conditions such as neurodegenerative diseases and inflammation. An important tool for the analysis of cellular metabolism is the application of stable isotope labeled substrates, which allow for the tracing of atoms throughout metabolic networks. While such analyses yield very detailed information about intracellular fluxes, the determination of compartment specific fluxes is far more challenging. Most approaches for the deconvolution of compartmented metabolism use computational models whereas experimental methods are rare. Here, we developed an experimental setup based on selective permeabilization of the cytosolic membrane that allows for the administration of stable isotope labeled substrates directly to mitochondria. We demonstrate how this approach can be used to infer metabolic changes in mitochondria induced by either chemical or genetic perturbations and give an outlook on its potential applications.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleAnalysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalMetabolic engineeringen
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