Protein Secretion: Methods and Protocols

ISBN-13: 9781603271677 / Angielski / Twarda / 2010 / 426 str.

A. Economou; Anastassios Economou

Opis

Over the past thirty years, many elegant genetic and biochemical approaches have been combined in order to advance the study of protein secretion and the necessary navigation through cell membranes, yet, despite this progress, less than two hundred membrane protein structures are known, nowhere near the complete inventory that the discovered protein export systems suggest. In Protein Secretion: Methods and Protocols, leading experts in the field provide robust, well-established protocols to elucidate the multiplicity of tools that have been developed to study protein sorting, membrane targeting, transmembrane crossing, and secretion across multiple membranes. With examples involving both prokaryotic and eukaryotic organisms, the volume covers subjects ranging from bioinformatics and proteomics to fundamental enzymology and genetics to cell biology, structural analyses, and biophysics. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters contain introductions to their respective topics, lists of the key materials and reagents, step-by-step, readily reproducible protocols, and detailed notes on troubleshooting and avoiding known pitfalls. Comprehensive and dependable, Protein Secretion: Methods and Protocols focuses on well-characterized paradigms so that scientists studying a vast array of subjects from biochemistry and genetics to biotechnology and biopharmaceuticals can benefit and expand upon their vital research.

Spis treści

1. The Extraordinary Diversity of Bacterial Protein Secretion Mechanisms I. Barry Holland 2. In vitro and in vivo Approaches to Studying the Bacterial Signal Peptide Processing Peng Wang and Ross E. Dalbey 3. Membrane Insertion of Small Proteins Andreas Kuhn, Natalie Stiegler, and Anne-Kathrin Schubert 4. Membrane Protein Insertion in E. coli Jijun Yuan, Ross E. Dalbey, and Andreas Kuhn 5. Study of Polytopic Membrane Protein Topological Organization as a Function of Membrane Lipid Composition Mikhail Bogdanov, Philip N. Heacock, and William Dowhan 6. In vivo Analysis of Protein Translocation to the Escherichia coli Periplasm Dominique Belin 7. Sorting of Bacterial Lipoproteins to the Outer Membrane by the Lol System Shin-ichiro Narita and Hajime Tokuda 8. Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex Ilja Kusters, Geert van den Bogaart, Janny de Wit, Viktor Krasnikov, Bert Poolman, and Arnold Driessen 9. Purification and Reconstitution of the SecY Translocon in Nanodiscs Kush Dalal and Franck Duong 10. In vitro Assays to Analyze Translocation of the Model Secretory Preprotein Alkaline Phosphatase Giorgos Gouridis, Spyridoula Karamanou, Marina Koukaki, and Anastassios Economou 11. Characterization of Interactions between Proteins Using Site-Directed Spin Labeling and Electron Paramagnetic Resonance Spectroscopy Jennine M. Crane, Angela A. Lilly, and Linda L. Randall 12. Analysis of Tat Targeting Function and Twin-Arginine Signal Peptide Activity in Escherichia coli Tracy Palmer, Ben C. Berks, and Frank Sargent 13. Site-Specific Cross-Linking of in Vitro Synthesized E. coli Preproteins for Investigating Transmembrane Translocation Pathways Sascha Panahandeh and Matthias Müller 14. Tracking theSecretion of Fluorescently-Labeled Type III Effectors from Single Bacteria in Real Time Nandi Simpson, Laurent Audry, and Jost Enninga 15. Comparative Analysis of Cytoplasmic Membrane Proteomes of Escherichia coli Using 2D Blue Native/SDS-PAGE Susan Schlegel, Mirjam Klepsch, David Wickström, Samuel Wagner, and Jan-Willem de Gier 16. Using Hidden Markov Models to Discover New Protein Transport Machines Vladimir A. Likic, Pavel Dolezal, Nermin Celik, Michael Dagley, and Trevor Lithgow 17. Bioinformatics Predictions of Localization and Targeting Shruti Rastogi and Burkhard Rosta 18. The Chloroplast Protein Import Machinery: A Review Penelope Strittmatter, Jürgen Soll, and Bettina Bölter 19. Measurement of the Energetics and Protein Transport Across the Chloroplast Thylakoid Membrane Steven M. Theg 20. In Vitro Dissection of Protein Translocation into the Mammalian Endoplasmic Reticulum Ajay Sharma, Malaiyalam Mariappan, Suhila Appathurai, and Ramanujan S. Hegde 21. In Vitro Reconstitution of the Selection, Ubiquitination, and Membrane Extraction of a Polytopic ERAD Substrate Kunio Nakatsukasa and Jeffrey L. Brodsky 22. Studying the ArfGAP-Dependent Conformational Changes in SNAREs Fernanda Rodriguez and Anne Spang 23. Studying Endoplasmic Reticulum Function in vitro Using siRNA Cornelia M. Wilson and Stephen High 24. High-Quality Immunofluorescence of Cultured Cells Dibyendu Bhattacharyya, Adam T. Hammond, and Benjamin S. Glick 25. Trapping Oxidative Folding Intermediates During Translocation to the Intermembrane Space of Mitochondia: in Vivo and in Vitro Studies Dionisia P. Sideris and Kostas Tokatlidis 26. Native Techniques for Analysis of Mitochondrial Protein Import F.-Nora Vögtle, Oliver Schmidt, Agnieszka Chacinska, Nikolaus Pfanner, and Chris Meisinger

Blurb

Over the past thirty years, many elegant genetic and biochemical approaches have been combined in order to advance the study of protein secretion and the necessary navigation through cell membranes, yet, despite this progress, less than two hundred membrane protein structures are known, nowhere near the complete inventory that the discovered protein export systems suggest. In Protein Secretion: Methods and Protocols, leading experts in the field provide robust, well-established protocols to elucidate the multiplicity of tools that have been developed to study protein sorting, membrane targeting, transmembrane crossing, and secretion across multiple membranes. With examples involving both prokaryotic and eukaryotic organisms, the volume covers subjects ranging from bioinformatics and proteomics to fundamental enzymology and genetics to cell biology, structural analyses, and biophysics. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the key materials and reagents, step-by-step, readily reproducible protocols, and detailed notes on troubleshooting and avoiding known pitfalls.

Comprehensive and dependable, Protein Secretion: Methods and Protocols focuses on well-characterized paradigms so that scientists studying a vast array of subjects from biochemistry and genetics to biotechnology and biopharmaceuticals can benefit and expand upon their vital research.