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Proteomic analyses have proven to be an invaluable tool in obtaining high-throughput protein identification from low-abundance, complex biological samples. In Organelle Proteomics, detailed protocols provide step-by-step instructions to successfully study organelle proteomes by performing the purification of the various organelles present in eukaryotic cells, as well as by preparing certain sub-fractions of organelles. A series of chapters cover the whole analytical procedure of organelle characterization, from its purification starting with whole cells up to protein identification using mass spectrometry. Devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle, Organelle Proteomics allows scientists to gain a vital and important understanding of organelle study.
List of contents
Analysis of Organelles by On-Line Two-Dimensional Liquid Chromatography-Tandem Mass Spectrometry.- Analysis of Organelles by On-Line Two-Dimensional Liquid Chromatography-Tandem Mass Spectrometry.- Analysis of Organelles, Vesicles, and Protein Machineries.- Purification and Proteomic Analysis of Chloroplasts and their Sub-Organellar Compartments.- Proteomic Analysis of the Stacked Golgi Complex.- Purification of Saccharomyces cerevisiae Mitochondria by Zone Electrophoresis in a Free Flow Device.- Preparation of Respiratory Chain Complexes from Saccharomyces cerevisiae Wild-Type and Mutant Mitochondria.- Mitochondrial Proteomics.- Purification and Proteomic Analysis of the Mouse Liver Mitochondrial Inner Membrane.- Subcellular Fractionation and Proteomics of Nuclear Envelopes.- Purification and Proteomic Analysis of a Nuclear-Insoluble Protein Fraction.- Preparation Methods of Human Metaphase Chromosomes for their Proteome Analysis.- Purification and Proteomic Analysis of Plant Plasma Membranes.- Protocol to Enrich and Analyze Plasma Membrane Proteins from Frozen Tissues.- Proteomic Characterization of Membrane Protein Topology.- Free Flow Isoelectric Focusing.- Use of Gas-Phase Fractionation to Increase Protein Identifications.- Purification and Proteomic Analysis of Lysosomal Integral Membrane Proteins.- Affinity Purification of Soluble Lysosomal Proteins for Mass Spectrometric Identification.- Purification and Proteomic Analysis of Synaptic Vesicles.- Purification and Proteomics Analysis of Pancreatic Zymogen Granule Membranes.- Isolation and Proteomic Analysis of Chlamydomonas Centrioles.- Purification and Proteomic Analysis of 20S Proteasomes from Human Cells.- Characterization of E. coli Ribosomal Particles.- Cellular, Genetic and Molecular Mechanisms of PlantEmbryogenesis.- Assessment of Organelle Purity Using Antibodies and Specific Assays.- Identifying Bona Fide Components of an Organelle by Isotope-Coded Labeling of Subcellular Fractions.- Determination of Genuine Residents of Plant Endomembrane Organelles using Isotope Tagging and Multivariate Statistics.- Quantitative Proteomic Analysis to Profile Dynamic Changes in the Spatial Distribution of Cellular Proteins.- Use of Transcriptomic Data to Support Organelle Proteomic Analysis.
Summary
Proteomic analyses have proven to be an invaluable tool in obtaining high-throughput protein identification from low-abundance, complex biological samples. In Organelle Proteomics, detailed protocols provide step-by-step instructions to successfully study organelle proteomes by performing the purification of the various organelles present in eukaryotic cells, as well as by preparing certain sub-fractions of organelles. A series of chapters cover the whole analytical procedure of organelle characterization, from its purification starting with whole cells up to protein identification using mass spectrometry. Devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle, Organelle Proteomics allows scientists to gain a vital and important understanding of organelle study.
