article · 20/10/2021
Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts
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.
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Prola, A., Vandestienne, A., Baroudi, N., Joubert, F., Tiret, L., & Pilot-Storck, F. (2021). Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts. Bio Protoc, 11(20):e4201. https://doi.org/https://doi.org/10.21769/bioprotoc.4201@article{Prola2021_363,
author = {Prola, Alexandre and Vandestienne, Aymeline and Baroudi, Nabil and Joubert, Frederic and Tiret, Laurent and Pilot-Storck, Fanny},
year = {2021},
month = {10},
title = {Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts},
journal = {Bio Protoc},
volume = {11(20):e4201},
abstract = {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.},
doi = {https://doi.org/10.21769/bioprotoc.4201},
}TY - JOUR
AU - Prola, Alexandre
AU - Vandestienne, Aymeline
AU - Baroudi, Nabil
AU - Joubert, Frederic
AU - Tiret, Laurent
AU - Pilot-Storck, Fanny
PY - 2021
DA - 2021/10/20
TI - Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts
JO - Bio Protoc
VL - 11(20):e4201
AB - 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.
DO - https://doi.org/10.21769/bioprotoc.4201
ER -