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Part 1 - Isolation 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
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 β-oxidation metabolites
Michael J. Bennett, Sheng Feng and Ann Saada (Reisch)
6. Biochemical assays of respiratory chain complex activity
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
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
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)
A. Basic properties of mitochondria
B. Inhibitors of the respiratory chain and of ATP synthesis
Nanette Orme-Johnson and Eric A. Schon
C. Linearized maps of selected mitochondrial genomes
D. Mitochondrial genetic codes in various organisms
E. Gene products present in mitochondria of yeast and animal cells
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.
- Provides a step-by-step "cookbook" presentation as written by leaders in the field
- Covers longstanding methods that have shaped the field
- Includes the newest technologies and methods
Research scientists in cell biology, developmental biology, and genetics. Academic and research libraries, pharmaceutical and biotechnology companies, and private research facilities
- No. of pages:
- © Academic Press 2020
- 13th March 2020
- Academic Press
- eBook ISBN:
- Hardcover ISBN:
Liza A. Pon, PhD, is a Professor of Pathology and Cell Biology and the Institute of Human Nutrition, and Director of the Confocal and Specialized Microscopy Shared Resource at Columbia University. As a PhD student at Tufts University, she carried out the first characterization of a protein now known as StaR, steroidogenic acute regulatory protein, which mediates transfer of cholesterol within mitochondria during steroid hormone biosynthesis. As a post-doctoral scientist with Gottfried (Jeff) Schatz at the University of Basel, she studied the mechanism of import of proteins into mitochondria. Dr. Pon joined Columbia University in 1990 and has been there ever since. Her research focuses on mitochondrial motility, dynamics, quality control and interactions with other organelles, and how these processes affect cell cycle progression, cellular fitness and lifespan. Recent studies in the laboratory focus on mechanisms for detection and removal of unfolded and damaged proteins not just in mitochondria but also in the ER in models of aging and disease.
Department of Pathology and Cell Biology and Institute of Human Nutrition, Columbia University
Eric A. Schon, PhD, is the Lewis P. Rowland Professor of Neurology (in Genetics and Development) at Columbia University. After graduating Columbia University with a B.S. in Chemical Engineering, he spent 10 years as a Technical Brand Manager for the Procter & Gamble Company in Cincinnati, OH. After leaving P&G, he received a PhD in Biological Chemistry from the University of Cincinnati, followed by a postdoctoral fellowship at the Harvard Medical School. He moved to Columbia as an Associate Research Scientist in 1984, and has been there ever since. Dr. Schon's laboratory studies the molecular genetics of neurological and neuromuscular diseases, with particular focus on mitochondrial disorders. The research has two principal goals: (1) to use the tools of molecular and cell biology in order to gain insight into the etiology, pathogenesis, and treatment of these devastating diseases, and (2) to build on this knowledge in order to ask more fundamental biological questions relating to nuclear-mitochondrial communication, mitochondrial biogenesis, and mtDNA plasticity. Most recently, the laboratory has become interested in understanding the structural and functional relationships between mitochondria and the endoplasmic reticulum in the pathogenesis of Alzheimer disease.
Department of Neurology (in Genetics and Development), Columbia University
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