Format:
Online-Ressource (LIV, 750 S.)
Edition:
Online-Ausg. 2011 Electronic reproduction; Available via World Wide Web
ISBN:
9780123745484
Series Statement:
Methods in enzymology 451
Content:
This is the companion volume to Daniel Klionsky's Autophagy: Lower Eukaryotes, which features the basic methods in autophagy covering yeasts and alternative fungi (aspergillus, podospora, magnaporthe). Klionsky is one of the leading authorities in the field. He is the editor-in-chief of Autophagy. The November 2007 issue of Nature Reviewshighlighted his article, Autophagy: from phenomenology to molecular understanding in less than a decade. He is currently editing guidelines for the field, with 230 contributing authors, that will publish in Autophagy. Particularly in times of stress, like starvation and disease, higher organisms have an internal mechanism in their cells for chewing up and recycling parts of themselves. The process of internal house-cleaning in the cell is called autophagy - literally self-eating. Breakthroughs in understanding the molecular basis of autophagy came after the cloning of ATG1 (autophagy-related gene 1) in yeast. (To date, 30 additional yeast genes have been identified.) These ATG genes in yeast were the stepping stones to the explosion of research into the molecular analysis of autophagy in higher eukaryotes. In the future, this research will help to design clinical approaches that can turn on autophagy and halt tumor growth. *Establishes the functional roles of specific cellular proteins in selective and nonselective autophagy in mammalian cells which aids researchers in determining why autophagy is shut down in neoplastia (growth of abnormal tissue mass) and turned on during bacterial invasion. *Includes methods to evaluate the role of autophagy in the drug-induced cell death of cancer cells in culture, which helps researchers design clinical approaches that can turn on autophagy and halt tumor growth. *Covers higher eukaryotes including lifespan in C. elegans to marine organisims and bridging into the clinical aspects, including autophagy in chronic myelogenous leukemia (CML is one of four types of leukaemia), lung cancer, prostate cancer, and cardiac cells.
Note:
Includes bibliographical references and indexes
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Front Cover; Autophagy: Lower Eukaryotes and Non-Mammalian Systems, Part A; Copyright Page; Contents; Contributors; Preface; Volumes in Series; Chapter 1: Biochemical Methods to Monitor Autophagy-Related Processes in Yeast; 1. Introduction; 3. Assays to Monitor Pexophagy; 5. Additional Methods; Chapter 2: Viability Assays to Monitor Yeast Autophagy; 2. Estimation of Yeast Viability with Phloxine B; Chapter 3: The Quantitative Pho8Delta60 Assay of Nonspecific Autophagy; 2. The Pho8Delta60 Assay; 3. Interpretation of the Results
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Chapter 4: Fluorescence Microscopy-Based Assays for Monitoring Yeast Atg Protein Trafficking1. Introduction; 3. Construction of the prApe1 and Atg8 Fluorescent Fusions; 4. Visualization of the Fluorescence Signals; 5. Atg9 Trafficking and the TAKA Assay; 6. Creation of the Strains for the Analysis of Atg9 Trafficking; References; Chapter 5: Measuring Macroautophagy in S. cerevisiae: Autophagic Body Accumulation and Total Protein Turnover; 2. Qualitative Measurement of Macroautophagy in S. Cerevisiae; 3. Quantitative Measurement of Macroautophagy in S. Cerevisiae
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Chapter 6: Aminopeptidase I Enzymatic Activity1. Introduction; 2. Methods to Measure Leucine-Aminopeptidase Activity; References; Chapter 7: Monitoring Autophagy in Yeast using FM 4-64 Fluorescence; 2. Molecular Properties and Endosomal-pathway Tracking Behavior of the FM 4-64 Fluorophore; Chapter 8: Monitoring Mitophagy in Yeast; 1. Introduction; 2. Visualization of Mitochondrial Sequestration within the Vacuole by Fluorescence Microscopy; References; Chapter 9: Monitoring Organelle Turnover in Yeast Using Fluorescent Protein Tags; 1. Selective Organelle Degradation by Autophagy in Yeast
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2. Principle of the Assay and General Considerations3. Targeting of Fluorescent Proteins (FPs) to Different Organelles; 4. Materials and Methods; 5. Analysis and Typical Images of Organellar Turnover; 6. Summary and Perspectives; Chapter 10: Electron Microscopy in Yeast; 1. Introduction; 2. Morphological Examination of Membrane Dynamics in Autophagy and the Cvt Pathway; 5. Morphometric Analysis of Autophagy-Related Structures; Acknowledgments; References; Chapter 11: Cell-Free Reconstitution of Microautophagy in Yeast; 2. Why Reconstitute Microautophagy In Vitro with Yeast Vacuoles?
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3. Methods for Reconstituting Microautophagy with Isolated VacuolesChapter 12: Autophagy in Wine Making; 1. Introduction; 3. Detection of Autophagy in Wine-making Using Industrial Yeast Strains; Chapter 13: Purification and in Vitro Analysis of Yeast Vacuoles; Acknowledgments; References; Chapter 14: Pexophagy in Hansenula polymorpha; 1. Introduction; 3. Cultivation of H. Polymorpha and Induction of Pexophagy; 4. Analysis of Peroxisome Degradation; 5. Concluding Remarks; Chapter 15: Pexophagy in Pichia pastoris; 1. Introduction: Use of Pichia Pastoris as a Model Organism to Study Pexophagy
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2. Culture Methods to Induce Micropexophagy or Macropexophagy in P. Pastoris
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Electronic reproduction; Available via World Wide Web
In:
Pt. A
Additional Edition:
Erscheint auch als Druck-Ausgabe Autophagy: lower eukaryotes and non-mammalian systems ; Pt. A Amsterdam [u.a.] : Elsevier, Acad. Press, 2008 ISBN 9780123745484
Language:
English
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