Methods in Cell Biology Volume 155 provides an update on the step-by-step "how-to" methods to study mitochondrial structure, function and biogenesis contained in the first two editions. As in the previous editions, biochemical, cell biological, and genetic approaches are presented along with sample results, interpretations, and pitfalls for each method. New chapters in this update include Isolation of Mitochondria and Analysis of Mitochondrial Compartments, Isolation of Mitochondria from Animal Cells and Yeast, Isolation and Characterization of Mitochondria-Associated ER Membranes, Import of Proteins into Mitochondria, Proximity Labeling Methods to Assess Protein-Protein Interactions in Yeast Mitochondria, and more.
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Table of Contents
Part 1Isolation of Mitochondria and Analysis of Mitochondrial Compartments 1. Isolation of mitochondria from animal cells and yeast Francesco Pallotti and Liza Pon 2. Isolation and characterization of mitochondria-associated ER membranes Estela Area-Gomez 3. Import of proteins into mitochondria Nikolaus Pfanner and Thomas Becker 4. Proximity labeling methods to assess protein-protein interactions in yeast mitochondria Jie-Ning Yang and Liza Pon
Part 2
Assays for Mitochondrial Metabolic Activities 5. Biochemical assays of TCA cycle and ?-oxidation metabolites Michael J. Bennett, Sheng Feng and Ann Saada (Reisch) 6. Biochemical assays of respiratory chain complex activity Robert Taylor 7. Assessing mitochondrial bioenergetics in whole cells and isolated organelles by microplate respirometry and polarography Erich Gnaiger and Orian Shirihai 8. Analyzing electron transport chain supercomplexes Jose Antonio Enriquez 9. Assays of mitochondrial ATP synthesis in animal cells and tissues Paolo Pinton and Hiromi Imamura 10. Visualization of mitochondrial membrane potential in mammalian cells Noemi Esteras, Merel Adjobo-Hermans, Andray Abramov and Werner JH Koopman 11. Visualization of mitochondrial function in living cells using biosensors (SoNAR, ro-GFP, DiOC6/Tom70 mCherry; DHE; Mitosox) Yi Yang and Liza Pon
Part 3
Assays for Other Mitochondrial Activities 12. Histochemical methods to measure respiratory chain activity and visualize mitochondria and mitochondrial gene products Kurenai Tanji and Sandra Franco-Iborra 13. Analysis of mitochondrial phospholipids and cardiolipin Michael Schlame and Estela Area-Gomez 14. Biosensors for Detection of Calcium and pH Diego Destefani 15. Measurement of membrane permeability and the mitochondrial permeability transition Paolo Bernardi and Michela Carraro
Part 4
Mitochondrial Genes and Gene Expression 16. Detection of mtDNA mutations Ali Naini, Sara Shanske, Robert Gilkerson and Jiuhong Pang 17. Visualization of mitochondrial DNA replication Agnel Sfeir 18. Cybrid technology Carlos Moraes and Sandra Bacman 19. Manipulation of mitochondrial genes and mtDNA heteroplasmy Carlos Moraes, Sandra Bacman and Michal Minczuk
Part?5
Assays for Mitochondrial Dynamics 20. Assays for mitochondrial fusion and fission Laura Lackner, Jason Lee, Jason Mears and Suzanne Hoppins 21. Methods to visualize mitochondria and mitochondrial motility Liza Pon, Theresa Swayne, Ryo Higuchi-Sanabria and Pin-Chao Liao 22. In vitro and in vivo detection of mitophagy in animal cells, mice, and yeast (Keima; red/green mitophagosomes)
Appendices A. Basic properties of mitochondria Luis Garc?a-Rodr?guez B. Inhibitors of the respiratory chain and of ATP synthesis Nanette Orme-Johnson and Eric A. Schon C. Linearized maps of selected mitochondrial genomes Eric Schon D. Mitochondrial genetic codes in various organisms Eric Schon E. Gene products present in mitochondria of yeast and animal cells Eric Schon
