Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts.
Acridine Orange 10-Nonyl Bromide
Cardiolipin
Mitotracker
NAO
Phosphatidylcholine
TopFluor CL
Vesicles
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
20 Oct 2021
20 Oct 2021
Historique:
received:
18
05
2021
revised:
28
07
2021
accepted:
06
08
2021
entrez:
11
11
2021
pubmed:
12
11
2021
medline:
12
11
2021
Statut:
epublish
Résumé
The efficient ATP production in mitochondria relies on the highly specific organization of its double membrane. Notably, the inner mitochondrial membrane (IMM) displays a massive surface extension through its folding into cristae, along which concentrate respiratory complexes and oligomers of the ATP synthase. Evidence has accumulated to highlight the importance of a specific phospholipid composition of the IMM to support mitochondrial oxidative phosphorylation. Contribution of specific phospholipids to mitochondrial ATP production is classically studied by modulating the activity of enzymes involved in their synthesis, but the interconnection of phospholipid synthesis pathways often impedes the determination of the precise role of each phospholipid. Here, we describe a protocol to specifically enrich mitochondrial membranes with cardiolipin or phosphatidylcholine, as well as a fluorescence-based method to quantify phospholipid enrichment. This method, based on the fusion of lipid vesicles with isolated mitochondria, may further allow a precise evaluation of phospholipid contribution to mitochondrial functions.
Identifiants
pubmed: 34761073
doi: 10.21769/BioProtoc.4201
pii: e4201
pmc: PMC8554811
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4201Informations de copyright
Copyright © 2021 The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare that they have no conflicts of interest or competing interests.
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