
Mitochondrial Translocases Part A
- 1st Edition, Volume 706 - October 24, 2024
- Imprint: Academic Press
- Editor: Nils Wiedemann
- Language: English
- Hardback ISBN:9 7 8 - 0 - 4 4 3 - 2 9 3 3 0 - 6
- eBook ISBN:9 7 8 - 0 - 4 4 3 - 2 9 3 3 1 - 3
Mitochondrial Translocases Part A series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an intern… Read more

Purchase options

Institutional subscription on ScienceDirect
Request a sales quoteMitochondrial Translocases Part A series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.
- Provides the latest information on biological research
- Offers outstanding and original reviews on a range of biological research topics
- Serves as an indispensable reference for researchers and students alike
Provides invaluable information on the fast-moving field of biological research that includes outstanding original reviews on a variety of topics
- Title of Book
- Cover image
- Title page
- Table of Contents
- Series Page
- Copyright
- Contributors
- Preface
- Section 1 Isolation of mitochondria
- Chapter One: Isolation of yeast mitochondria by differential centrifugation
- Abstract
- 1 Introduction
- 2 Before you begin
- 3 Key resources table
- 4 Materials and equipment
- 5 Step-by-step method details
- 6 Expected outcomes for isolated yeast mitochondria
- 7 Advantages
- 8 Limitations
- 9 Optimization and troubleshooting
- 10 Safety considerations and standards
- Acknowledgments
- References
- Chapter Two: Isolation of yeast mitochondria by affinity purification using magnetic beads
- Abstract
- 1 Introduction
- 2 Before you begin
- 3 Key resources table
- 4 Materials and equipment
- 5 Step-by-step method details
- 6 Expected outcomes
- 7 Advantages
- 8 Limitations
- 9 Optimization and troubleshooting
- 10 Alternative methods/procedures
- 11 Summary/Conclusion
- Acknowledgments
- References
- Chapter Three: A modified procedure for separating yeast peroxisomes from mitochondria
- Abstract
- 1 Introduction
- 2 Yeast growth conditions
- 3 Preparation of yeast spheroplasts
- 4 Preparation of yeast cell lysates and isolation of crude organelles
- 5 Discontinuous gradient to separate mitochondria and peroxisomes
- Acknowledgments
- References
- Section 2 Analysis of mitochondrial proteins
- Chapter Four: Subcellular fractionation by differential centrifugation for mitochondrial studies
- Abstract
- 1 Introduction
- 2 General considerations
- 3 Experimental design
- 4 Subcellular fractionation of yeast cells by differential centrifugation
- 5 Conclusion and summary
- Acknowledgments
- References
- Chapter Five: Monitoring mitochondrial localization of dual localized proteins using a Bi-Genomic Mitochondrial-Split-GFP
- Abstract
- 1 Introduction
- 2 Materials
- 3 Methods
- Acknowledgments
- References
- Chapter Six: Analysis of mitochondrial biogenesis and protein localization by genetic screens and automated imaging
- Abstract
- 1 Introduction to yeast genetics
- 2 Systematic mutant collections: past and present
- 3 Creation of “tailor made” collections
- 4 Interrogating collections
- 5 Discovery of mitochondrial biology using systematic mutant libraries
- 6 Perspective
- Acknowledgments
- References
- Chapter Seven: Methods to monitor mitochondrial disulfide bonds
- Abstract
- Abbreviations
- 1 Introduction
- 2 Strategies to monitor mitochondrial disulfide bonds
- 3 Direct and indirect thiol trap assays
- 4 Strategies to identify MIA40 substrates
- 5 Import of Mia40 substrates
- Acknowledgment
- References
- Section 3 Prediction, analysis and processing of mitochondrial targeting signals
- Chapter Eight: Prediction of mitochondrial targeting signals and their cleavage sites
- Abstract
- 1 Introduction
- 2 Use and installation of MTS prediction tools
- 3 Resources for MTS dataset construction
- 4 Retrospective case study
- 5 Summary of computational methods used for MTS prediction
- 6 Regarding predictions on “atypical” proteins
- 7 Future directions of MTS and similar prediction tasks
- 8 Summary
- Acknowledgments
- References
- Chapter Nine: Monitoring mitochondrial precursor processing and presequence peptide degradation
- Abstract
- 1 Introduction
- 2 Analysis of MPP activity in vitro
- 3 Analysis of precursor processing and presequence degradation in mitochondrial soluble extracts
- Acknowledgments
- References
- Chapter Ten: Analysis of mitochondrial targeting signal cleavage and protein processing by mass spectrometry
- Abstract
- 1 Introduction
- 2 Enrichment of N-terminal peptides
- 3 N-terminome data analysis and termini annotation
- 4 Expected outcomes
- 5 Alternative methods
- Acknowledgments
- References
- Chapter Eleven: Analysis of targeting signals for mitochondrial intermembrane space import
- Abstract
- Abbreviations
- 1 Introduction
- 2 Precursors and import assays
- 3 Post import analysis and sub-fractionation
- 4 Overview and conclusions
- Acknowledgments
- References
- Chapter Twelve: Analysis and prediction of internal mitochondrial targeting signals
- Abstract
- 1 Introduction
- 2 Mitochondrial protein import: Routes and targeting sequences
- 3 Internal targeting sequences: The challenge of iMTS identification
- 4 Machine learning as a generic approach for iMTS identification
- 5 iMTS prediction with TargetP
- 6 Facilitating iMTS identification using the predictor iMLP
- 7 Limitations
- 8 Optimization and troubleshooting
- 9 Concluding remark
- Acknowledgments
- References
- Section 4 Mitochondrial precursor proteins in the cytosol
- Chapter Thirteen: Monitoring and analysis of mitochondrial precursor protein aggregates in the cytosol
- Abstract
- 1 Introduction
- 2 Monitoring and analysis of mitochondrial precursor protein aggregates in the cytosol of yeast
- 3 Detection of mitochondrial precursor protein aggregates via immunofluorescence microscopy
- 4 Studying the effects of mitochondrial precursor aggregation on cellular proteostasis via fluorescence microscopy
- 5 Monitoring and analysis of mitochondrial precursor protein aggregates in the cytosol of human cells
- 6 Overview and future perspectives
- Acknowledgments
- References
- Chapter Fourteen: In vitro translation in yeast extract to study interactions with cytosolic chaperones
- Abstract
- 1 Introduction
- 2 Yeast cells used for in vitro translation
- 3 Yeast extract preparation for in vitro translation
- 4 Preparation of a plasmid encoding the protein of interest for translation in yeast extract
- 5 In vitro transcription of the gene of interest
- 6 In vitro translation reaction in yeast extract
- 7 Pull down assay
- Acknowledgments
- References
- Section 5 Mitochondrial protein translocation
- Chapter Fifteen: In vitro import of mitochondrial precursor proteins into yeast mitochondria
- Abstract
- 1 Introduction
- 2 In vitro synthesis of mitochondrial precursor proteins
- 3 In vitro import into isolated yeast mitochondria
- Acknowledgments
- References
- Chapter Sixteen: Monitoring the in vitro import and assembly of mitochondrial precursor proteins into mammalian mitochondria
- Abstract
- 1 Introduction
- 2 In vitro synthesis of 35S-radiolabelled precursor proteins
- 3 Mitochondrial isolation
- 4 In vitro import assay
- 5 Monitoring assembly into protein complexes
- 6 Visualising in vitro import reactions
- 7 Concluding remarks
- Acknowledgements
- References
- Chapter Seventeen: Import of mitochondrial precursor proteins into mitochondria of semi-intact yeast cells
- Abstract
- 1 Introduction
- 2 Mitochondrial protein import: routes and targeting sequences
- 3 Semi-intact cells
- 4 Import of proteins into semi-intact cells
- 5 Notes
- 6 General considerations
- Acknowledgments
- References
- Chapter Eighteen: The MitoLuc assay for the analysis of the mechanism of mitochondrial protein import
- Abstract
- 1 Introduction
- 2 Methods
- References
- Chapter Nineteen: Monitoring mitochondrial protein import by live cell imaging
- Abstract
- 1 Introduction
- 2 Equipment and materials
- 3 Protocol
- 4 Notes
- Acknowledgment
- References
- Chapter Twenty: Quantifying mitochondrial protein import by mePRODmt proteomics
- Abstract
- 1 Introduction
- 2 Materials
- 3 Methods
- Acknowledgments
- References
- Chapter Twenty One: Methods to analyze mitochondrial protein translocation in plant mitochondria
- Abstract
- 1 Introduction
- 2 Plants and tissues
- 3 Dual-targeting verse mis-targeting
- 4 The import of precursor proteins into isolated plant mitochondria
- 5 Protein import assays using freshly isolated mitochondria
- 6 In vivo protein uptake studies with fluorescent protein tagging
- References
- Section 6 Mitochondrial gene expression and translation
- Chapter Twenty Two: In organello silencing of mitochondrial gene expression
- Abstract
- 1 Introduction
- 2 Protein carrier preparation
- 3 HEK293T mitochondrial isolation
- 4 In organello silencing of mitochondrial gene expression
- 5 Conclusion
- 6 Troubleshooting
- References
- Chapter Twenty Three: Assays to monitor mitochondrial translation
- Abstract
- 1 Introduction
- 2 Before you begin
- 3 Materials and equipment
- 4 Step-by-step method details
- 5 Expected outcomes
- 6 Quantification and statistical analysis
- 7 Advantages
- 8 Limitations
- 9 Optimization and troubleshooting
- 10 Safety considerations and standards
- References
- Chapter Twenty Four: Analysis of mitochondrial translation using click chemistry
- Abstract
- Abbreviations
- 1 Introduction
- 2 Cell culture
- 3 L-Homopropargylglycine (HPG) incorporation in living cells
- 4 HPG click chemistry in isolated mitochondria
- 5 HPG click chemistry for microscopy imaging
- 6 High resolution microscopy
- 7 Trouble shooting
- 8 Conclusions and outlook
- Acknowledgments
- References
- Edition: 1
- Volume: 706
- Published: October 24, 2024
- Imprint: Academic Press
- No. of pages: 576
- Language: English
- Hardback ISBN: 9780443293306
- eBook ISBN: 9780443293313
Read Mitochondrial Translocases Part A on ScienceDirect