Protein Targeting Protocols (eBook)

Mark van der Giezen (Herausgeber)

eBook Download: PDF | EPUB

2008 | 1. Auflage
495 Seiten
Humana Press (Verlag)
978-1-59745-466-7 (ISBN)

Infos

Currently not available, will follow before Dec 30.

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In this highly anticipated update of the extremely successful Protein Targeting Protocols, experts from around the world provide the latest protocols on for isolating different organelles and the localization of particular proteins using a variety of methods such as light, confocal, and electron microscopy. Unlike the first edition, however, this volume places emphasis on protein targeting of cellular compartments in both prokaryotic and eukaryotic systems.Authors provide state-of-the-art information on the fast moving fields of import machinery of mitochondria and plastids, as well as a detailed protocol on the movement of protein complexes in bacterial membranes using fluorescent recovery after photobleaching. Bacterial protein targeting protocols using the Sec-system, type-V secretion apparatus, and the Tat-pathway are described, as is a periplasmic targeting protocol. Chapters also include bioinformatics methods to guide readers through the ever-increasing number of in silico tools currently available. Editor Mark van der Giezen has succeeded in including targeting protocols from different systems, including animal, plant, fungal, and protist models, providing insights into the intricate challenges presented by these diverse systems.Authoritative and state-of-the-art, Protein Targeting Protocols, Second Edition, provides useful techniques that will aid scientists working with a variety of organisms.

Inhalt

Preface 6
Contents 8
Contributors 11
Bacterial Systems 16
Localization and Mobility of Bacterial Proteins by Confocal Microscopy and Fluorescence Recovery After Photobleaching 17
Membrane Protein Insertion and Secretion in Bacteria 30
Identification of Autotransporter Proteins Secreted by Type V Secretion Systems in Gram- Negative Bacteria 45
Periplasmic Targeting of Recombinant Proteins in Escherichia coli 59
In Vitro Analysis of the Bacterial Twin- Arginine- Dependent Protein Export 74
Eukaryotic Systems 91
Nuclear Import Properties of the Sex-Determining Factor SRY 92
The Mitochondrial Machinery for Import of Precursor Proteins 107
A Genetic Method to Identify Mitochondrial Proteins in Living Mammalian Cells 126
In Vitro and In Vivo Methods to Study Protein Import Into Plant Mitochondria 138
Protein Targeting to Mitochondria of Saccharomyces cerevisiae and Neurospora crassa 158
A Mitosome Purification Protocol Based on Percoll Density Gradients and Its Use in Validating the Mitosomal Nature of Entamoeba histolytica Mitochondrial Hsp70 174
The Chloroplast Protein Import Machinery: A Review 185
Import of Plastid Precursor Proteins Into Pea Chloroplasts 200
Protein Targeting in “Secondary” or “ Complex” Chloroplasts 212
Protein Trafficking to the Complex Chloroplasts of Euglena 223
Fluorescent Protein Fusions for Protein Localization in Plants 242
Genetic Transformation 259
Identification of Proteins Targeted Into the Endoplasmic Reticulum by cDNA Library Screening 270
Trafficking Through the Early Secretory Pathway of Mammalian Cells 282
Studying Protein Export From the Endoplasmic Reticulum in Plants 298
Imaging the Golgi Apparatus in Living Mitotic Cells 310
Evaluating Yeast Biosynthetic Vacuolar Transport 330
Targeting to Lysosomes in Mammalian Cells 339
Monitoring Autophagy in Yeast 362
Protein Targeting to Yeast Peroxisomes 371
Expression, Localization, and Topology of Fluorescently Tagged Plasma Membrane-Targeted Transmembrane Proteins 390
General Methods 401
Intracellular Protein Localization by Immunoelectron Microscopy 402
Separation of Proteins by Blue Native Electrophoresis 412
Computational Prediction of Subcellular Localization 423
Phylogenetic Analysis to Uncover Organellar Origins of Nuclear- Encoded Genes 461
Index 483

Produkt-Details

• Erscheint lt. Verlag: 1.4.2008
• Sprache: englisch
• Themenwelt: Studium ► 1. Studienabschnitt (Vorklinik) ► Biochemie / Molekularbiologie
• Themenwelt: Naturwissenschaften ► Biologie ► Biochemie
• Themenwelt: Technik
• ISBN-10: 1-59745-466-4 / 1597454664
• ISBN-13: 978-1-59745-466-7 / 9781597454667