This volume explores numerous techniques for the genetic, molecular, biochemical, and structural examination of BCL-2 family proteins and their interactions. The chapters in this book cover topics such as the relevance of BCL-2 proteins in health and disease; evaluating cellular dependencies to specific BCL-2 family proteins; flow-cytometry-based methods for measuring BCL-2 proteins and mitochondrial-based cell death; measuring activity and interactions of BCL-2 family proteins in the presence of mitochondria, artificial membranes or yeast; conformational activation and oligomerization of pro-apoptotic proteins BAX and BAK leading to cytochrome c release and apoptosis; structural and biophysical studies in solution and lipid vesicles using nuclear magnetic resonance, cryo-electron microscopy, fluorescence microscopy and electron paramagnetic resonance. Written in the highly successful
Methods in Molecular Biology
series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough,
BCL-2 Family Proteins: Methods and Protocols
is a valuable resource to inspire and encourage novice and established scientists to further their research and make new discoveries in this exciting field.
Reihe
Sprache
Verlagsort
Verlagsgruppe
Illustrationen
66
30 s/w Abbildungen, 66 farbige Abbildungen
XI, 360 p. 96 illus., 66 illus. in color.
Dateigröße
ISBN-13
978-1-4939-8861-7 (9781493988617)
DOI
10.1007/978-1-4939-8861-7
Schweitzer Klassifikation
Overview of BCL-2 Family Proteins and Therapeutic Potentials.- Database and Bioinformatic Analysis of BCL-2 Family Proteins and BH3-Only Proteins.- Application of Mito-Priming to Generate BCL-2 Addicted Cells.- BH3 Profiling: A Functional Assay to Measure Apoptotic Priming and Dependencies.- Flow Cytometry-Based Detection and Analysis of BCL-2 Family Proteins and Mitochondrial Outer Membrane Permeabilization (MOMP).- Investigating BCL-2 Family Protein Interactions in Yeast.- Liposomal Permeabilization Assay to Study the Functional Interaction of the BCL-2 Family.- Mitochondrial Isolation, Gel Filtration, and Real-Time Monitoring of MOMP.- Photocrosslinking Approach to Investigate Protein Interactions in the BCL-2 Family.- Assessment of Dynamic BCL-2 Protein Shuttling between Outer Mitochondrial Membrane and Cytosol.- Quantification of BCL-2 Family Members by Flow Cytometry.- Methods to Probe Calcium Regulation by BCL-2 FamilyMembers.- Methods to Probe Conformational Activation and Mitochondrial Activity of Proapoptotic BAK.- Probing BAK and BAX Activation and Pore Assembly with Cytochrome c Release, Limited Proteolysis and Oxidant-Induced Linkage.- BCL-2 Protein Family Interaction Analysis by Nuclear Magnetic Resonance Spectroscopy.- Reconstitution and Characterization of BCL-2 Family Proteins in Lipid Bilayer Nanodiscs.- Cryo-Electron Microscopy to Study Bax Pores and MOMP.- CW EPR and DEER Methods to Determine BCL-2 Family Protein Structure and Interactions: Application of Site-Directed Spin Labeling to BAK Apoptotic Pores.- Rapid Imaging of BCL-2 Family Interactions in Live Cells using FLIM-FRET.- Quantification of the Interactions between BCL-2 Proteins by Fluorescence Correlation Spectroscopy.- Isolation of Synthetic Antibodies against BCL-2-Associated X Protein (BAX).
