Organelle Proteomics

Proteomic analyses have proven to be an invaluable tool in obtaining high-throughput protein identification from low-abundance, complex biological samples. This is the first book that examines organelle proteomics in depth. 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. Also, devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle, the book allows scientists to gain a vital and important understanding of organelle study.

Collects time tested reproducible laboratory protocols including tricks and hints written by experts in the field Currently the only book that examines organelle proteomics in depth

Klappentext

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.

Inhalt
Analysis of Organelles by On-Line Two-Dimensional Liquid ChromatographyTandem Mass Spectrometry.- Analysis of Organelles by On-Line Two-Dimensional Liquid ChromatographyTandem 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.