Title Post-translational processing : a practical approach / edited by S.J. Higgins and B.D. Hames

Published Oxford ; New York : Oxford University Press, 1999

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Description 1 online resource (xx, 317 pages) : illustrations Series Practical approach series ; 203 Practical approach series ; 203. Contents Intro; Contents; List of Contributors; Abbreviations; 1. Sequence analysis of expressed proteins; 1. Introduction; 2. N-terminal sequence analysis; Automated sequencing; 3. Sample preparation; Sample preparation by SDS-PAGE; Sample preparation by HPLC; Other procedures for sample preparation; 4. N-terminal blocking; Protein fragmentation; 5. C-terminal sequencing; Chemical analysis of the C-terminus; Enzymic analysis of the C-terminus; 6. Mass spectrometric analysis of proteins; A brief guide to mass spectrometers; Molecular mass determination; Sequencing by mass spectrometry; References 2. Protein folding and import into organelles1. Introduction; 2. Preparation of preprotein substrates for in vitro import into organelles; Preparation of mRNA for in vitro translation; Synthesis of preproteins in the reticulocyte lysate; Synthesis of preproteins in wheat germ lysate; Synthesis of preproteins in yeast cytosol; Synthesis of preproteins in Escherichia coli; Dihydrofolate reductase as a model protein for import studies; 3. Import of proteins into mitochondria; Isolation of mitochondria for import studies; Import of preproteins into isolated mitochondria Generation of translocation intermediatesLocalization of imported proteins; 4. Import of proteins into microsomes (endoplasmic reticulum); Isolation of microsomes; Import of preproteins into microsomes; 5. Import of proteins into other organelles; 6. Analysis of protein import into organelles; Monitoring the association of proteins with membranes; Analysis of protein complexes involved in organelle import; Analysis of protein folding after import into organelles; Acknowledgements; References; 3. Analysis of protein phosphorylation; 1. Introduction 2. Investigating protein phosphorylation systems3. Phosphorylation of proteins in intact preparations; General considerations; Intact animals; Tissue slices; Isolated cells; 4. Phosphorylation of proteins in cell-free preparations; General considerations; Labelling and stimulation of cell-free preparations; 5. Analysis of phosphorylated proteins; General considerations; Quantification of phosphoproteins; Phosphoprotein separation; Protein isolation by immunomethods; Analysis of multisite phosphorylation; Phosphoamino acid analysis; Analysis of the state of phosphorylation 6. Analysis of protein kinasesGeneral considerations; Analysis of protein kinase activity in vitro; Analysis of specific protein kinases in vitro; Analysis of protein kinase activity in intact cells; Purification of protein kinases; 7. Phosphoprotein phosphatases; General considerations; P-Ser/P-Thr phosphoprotein phosphatases; Tyrosine-specific phosphoprotein phosphatases; References; 4. Protein glycosylation; 1. Introduction; Protein glycosylation; Oligosaccharide structures; Glycosylation pathways; Characterization of protein glycosylation; 2. Is my protein glycosylated? Summary Post - Translational Modification: A Practical Approach and its companion volume Protein Expression: A Practical Approach form the final part of the PAS mini-series on protein synthesis and processing. This volume begins with a chapter on protein sequencing followed by a chapter on protein folding and import into organelles. The next three chapters cover the three major forms of covalent modification: phosphorylation, glycosylation, and lipid modification. Proteolytic processing the is the next topic and the final two chapters are concerned with protein turnover in mammalian cells and yeast. This book is a comprehensive volume of the best current methodology and is designed to be used at the bench or away from the bench to gain insight into future experimental approached Bibliography Includes bibliographical references and index Notes Print version record Subject Post-translational modification. Protein Biosynthesis Peptide Biosynthesis SCIENCE -- Life Sciences -- Biochemistry. Post-translational modification Protéines -- Modifications posttraductionnelles. Form Electronic book Author Higgins, S. J. (Steve J.) Hames, B. D ISBN 0585483663 9780585483665 1280375760 9781280375767 9780191566035 0191566039

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