This book starts with a chapter by Dr. Edwin Romijn and Prof. John R. Yates III, who introduce the different analytical strategies developed and successfully utilized to study organelle proteomes, and detail the use of multidimensional liquid chromatography coupled to tandem mass spectrometry for peptide sample analysis. It is further composed of two main sections. First, detailed protocols are provided to perform the purification of the various organelles present in eukaryotic cells, as well as to prepare certain sub-fractions of organelles (chapters 2-22). In all cases, the samples are aimed to be analyzed by a mass spectrometry technique. While an exhaustive list of chapters covering all the proteomic analyses of organelles and organelle fractions was not conceivable, we nevertheless wanted to provide analysis examples reflecting the trend toward more specific purifications of organelle sub-fractions, which will allow reaching the more comprehensive and accurate characterization of the organelle. 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. INDICE: From the contents Analysis of organelles by on-line two-dimensional liquid chromatography – tandem mass spectrometry (2DLC-MS/MS).- Purificationand 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 (ZE-FFE).- Preparation of Respiratory Chain Complexes from Saccharomyces cerevisiae Wild Type and Mutant Mitochondria: Activity Measurement and Subunit Composition Analysis.- Mitochondrial proteomics: Analysis of a whole mitochondrial extract with two-dimensional electrophoresis.- Purification and proteomic analysis of the mouse liver mitochondrial inner membrane. - Subcellular Fractionationand Proteomics of Nuclear Envelopes.- Purification and proteomic analysis of the nuclear insoluble protein fraction.- Preparation Methods of Human Metaphase Chromosomes for Their Proteome Analysis.
- ISBN: 978-1-58829-779-2
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 490
- Fecha Publicación: 01/03/2008
- Nº Volúmenes: 1
- Idioma: Inglés