Molecular Characterization of Autophagic Responses Part A: Volume 587

Molecular Characterization of Autophagic Responses Part A: Volume 587

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Molecular Characterization of Autophagic Responses, Part A, presents a collection of methods for the qualitative and quantitative evaluation of virtually all the morphological, biochemical, and functional manifestations of autophagy, in vitro, ex vivo and in vivo, in organisms as distant as yeast and man.

Autophagy is an evolutionarily conserved mechanism for the lysosomal degradation of superfluous or dangerous cytoplasmic entities, and plays a critical role in the preservation of cellular and organismal homeostasis. Monitoring the biochemical processes that accompany autophagy is fundamental for understanding whether autophagic responses are efficient or dysfunctional.
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Product details

  • Hardback | 598 pages
  • 152 x 229 x 31.75mm | 1,090g
  • Academic Press Inc
  • San Diego, United States
  • English
  • 0128096756
  • 9780128096758

Table of contents

1. Correlative Live Cell and Super Resolution Imaging of Autophagosome Formation 2. Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy 3. The Use of DQ-BSA to Monitor the Turnover of Autophagy-Associated Cargo 4. Turnover of Lipidated LC3 and Autophagic Cargoes in Mammalian Cells 5. High-Throughput Quantification of GFP-LC3+ Dots by Automated Fluorescence Microscopy 6. Use of pHlurorin-mKate2-human LC3 to Monitor Autophagic Responses 7. Production of Human ATG Proteins for Lipidation Assays 8. Investigating Structure and Dynamics of Atg8 Family Proteins 9. Methods for Studying Interactions Between Atg8/LC3/GABARAP and LIR-Containing Proteins 10. Assessment of Posttranslational Modifications of ATG proteins 11. Tagged ATG8-Coding Constructs for the In Vitro and In Vivo Assessment of ATG4 Activity 12. Measurement of the Activity of the Atg4 Cysteine Proteases 13. Crystallographic Characterization of ATG Proteins and Their Interacting Partners 14. Dynamics of Atg5-Atg12-Atg16L1 Aggregation and Deaggregation 15. Fluorescent FYVE Chimeras to Quantify PtdIns3P Synthesis During Autophagy 16. Quantification of Phosphatidylinositol Phosphate Species in Purified Membranes 17. Mass Assays to Quantify Bioactive PtdIns3P and PtdIns5P During Autophagic Responses 18. Fluorescence-Based Assays to Analyse Phosphatidylinositol 5-Phosphate in Autophagy 19. Ultrastructural Characterization of Phagophores Using Electron Tomography on Cryoimmobilized and Freeze Substituted Samples 20. A Simple Cargo Sequestration Assay for Quantitative Measurement of Nonselective Autophagy in Cultured Cells 21. In Vitro Reconstitution of Autophagosome-Lysosome Fusion 22. In Vitro Reconstitution of Atg8 Conjugation and Deconjugation 23. Study of ULK1 Catalytic Activity and Its Regulation 24. Evaluating the mTOR Pathway in Physiological and Pharmacological Settings 25. Methods to Study the BECN1 Interactome in the Course of Autophagic Responses 26. In Vitro Characterization of VPS34 Lipid Kinase Inhibition by Small Molecules 27. Methods to Study Lysosomal AMPK Activation 28. Allosteric Modulation of AMPK Enzymatic Activity: In Vitro Characterization 29. Assessing the Catalytic Activity of Transglutaminases in the Context of Autophagic Responses
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Review Text

Praise for the Series: "Should be on the shelves of all libraries in the world as a whole collection." --Chemistry in Industry "The work most often consulted in the lab." -- Enzymologia "The Methods in Enzymology series represents the gold-standard." --Neuroscience
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Review quote

Praise for the Series: "Should be on the shelves of all libraries in the world as a whole collection." --Chemistry in Industry "The work most often consulted in the lab." --Enzymologia "The Methods in Enzymology series represents the gold-standard." --Neuroscience
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About Guido Kroemer

Lorenzo Galluzzi received his Ph.D. in 2008 from the University of Paris Sud/Paris XI (France), and now works as a research manager in the laboratory of Guido Kroemer. He is particularly fascinated by several aspects of mitochondrial cell death, autophagy, cancer cell metabolism and tumour immunology. He has published more than 270 articles in peer-reviewed scientific journals, and is currently the 6th and youngest of the 30 most-cited European cell biologists (relative to the period 2007-2013). Guido Kroemer got his M.D. in 1985 from the University of Innsbruck, Austria, and his Ph.D. in molecular biology in 1992 from the Autonomous University of Madrid, Spain. He is currently Professor at the Faculty of Medicine of the University of Paris Descartes/Paris V, Director of the INSERM research team 'Apoptosis, Cancer and Immunity', Director of the Metabolomics and Cell Biology platforms of the Gustave Roussy Cancer Campus, and Practitioner at the Hopital Europeen George Pompidou (Paris, France). He is also the Director of the Paris Alliance of Cancer Research Institutes (PACRI) and the Labex 'Immuno-Oncology'. Dr. Kroemer is best known for the discoveries that mitochondrial membrane permeabilization constitutes a decisive step in regulated cell death; that autophagy is a cytoprotective mechanism with lifespan-extending effects; and that anticancer therapies are successful only if they stimulate tumour-targeting immune responses. He is currently the most-cited cell biologist in Europe (relative to the period 2007-2013), and he has received the Descartes Prize of the European Union, the Carus Medal of the Leopoldina, the Dautrebande Prize of the Belgian Royal Academy of Medicine, the Leopold Griffuel Prize of the French Association for Cancer Research, the Mitjavile prize of the French National Academy of Medicine and a European Research Council Advanced Investigator Award. Jose Manuel Bravo-San Pedro graduated from the University of Extremadura (Caceres, Spain) in 2011, and now works as a post-doctoral fellow in the laboratory of Guido Kroemer. His main research interests encompass the molecular cross-talk between autophagy and regulated cell death, and the interconnections between cellular autophagic responses and organismal metabolism.
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