Bacterial protein secretion systems : methods and protocols

✍ Scribed by Cascales, Eric; Journet, Laure

• Publisher: Humana Press ; Springer
• Year: 2017
• Tongue: English
• Leaves: 521
• Series: Methods in molecular biology 1615
• Category: Library

✦ Synopsis

This volume details protocols that cover the broad arsenal of techniques used to study a secretion system from A to Z. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as reporters to follow effector transport. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Bacterial Protein Secretion Systems: Methods and Protocol aims to provide techniques that are not restricted to the study of secretion systems but are also of specific interest for any researcher interested on multi-protein complexes of the bacterial cell envelope. Read more...

Abstract: This volume details protocols that cover the broad arsenal of techniques used to study a secretion system from A to Z. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as reporters to follow effector transport. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Bacterial Protein Secretion Systems: Methods and Protocol aims to provide techniques that are not restricted to the study of secretion systems but are also of specific interest for any researcher interested on multi-protein complexes of the bacterial cell envelope

✦ Table of Contents

Front Matter ....Pages i-xv
Identification of Protein Secretion Systems in Bacterial Genomes Using MacSyFinder (Sophie S. Abby, Eduardo P. C. Rocha)....Pages 1-21
Protein Sorting Prediction (Henrik Nielsen)....Pages 23-57
Cell Fractionation (Melissa Petiti, Laetitia Houot, Denis Duché)....Pages 59-64
Defining Lipoprotein Localisation by Fluorescence Microscopy (Maria Guillermina Casabona, Mylène Robert-Genthon, Didier Grunwald, Ina Attrée)....Pages 65-74
Identification of Lipoproteins Using Globomycin and Radioactive Palmitate (Nienke Buddelmeijer)....Pages 75-80
Defining Membrane Protein Localization by Isopycnic Density Gradients (Rhys A. Dunstan, Iain D. Hay, Trevor Lithgow)....Pages 81-86
Cell Surface Exposure (Anna Konovalova)....Pages 87-95
Probing Inner Membrane Protein Topology by Proteolysis (Maxence S. Vincent, Eric Cascales)....Pages 97-103
Mapping of Membrane Protein Topology by Substituted Cysteine Accessibility Method (SCAM™) (Mikhail Bogdanov)....Pages 105-128
Defining Membrane Protein Topology Using pho-lac Reporter Fusions (Gouzel Karimova, Daniel Ladant)....Pages 129-142
In Vivo and In Vitro Protein–Peptidoglycan Interactions (Gang Li, S. Peter Howard)....Pages 143-149
Measure of Peptidoglycan Hydrolase Activity (Yoann G. Santin, Eric Cascales)....Pages 151-158
Protein–Protein Interaction: Bacterial Two-Hybrid (Gouzel Karimova, Emilie Gauliard, Marilyne Davi, Scot P. Ouellette, Daniel Ladant)....Pages 159-176
Protein–Protein Interactions: Yeast Two-Hybrid System (Jer-Sheng Lin, Erh-Min Lai)....Pages 177-187
Protein–Protein Interactions: Cytology Two-Hybrid (Krishnamohan Atmakuri)....Pages 189-197
Fusion Reporter Approaches to Monitoring Transmembrane Helix Interactions in Bacterial Membranes (Laureen Logger, Abdelrahim Zoued, Eric Cascales)....Pages 199-210
Protein–Protein Interactions: Co-Immunoprecipitation (Jer-Sheng Lin, Erh-Min Lai)....Pages 211-219
Protein–Protein Interaction: Tandem Affinity Purification in Bacteria (Julie P. M. Viala, Emmanuelle Bouveret)....Pages 221-232
Site-Directed and Time-Resolved Photocrosslinking in Cells Metabolically Labeled with Radioisotopes (Raffaele Ieva)....Pages 233-245
Protein–Protein Interactions: Pull-Down Assays (Arthur Louche, Suzana P. Salcedo, Sarah Bigot)....Pages 247-255
Protein–Protein Interactions: Surface Plasmon Resonance (Badreddine Douzi)....Pages 257-275
Assessing Energy-Dependent Protein Conformational Changes in the TonB System (Ray A. Larsen)....Pages 277-287
Defining Assembly Pathways by Fluorescence Microscopy (Abdelrahim Zoued, Andreas Diepold)....Pages 289-298
Large Complexes: Cloning Strategy, Production, and Purification (Eric Durand, Roland Lloubes)....Pages 299-309
Shearing and Enrichment of Extracellular Type IV Pili (Alba Katiria Gonzalez Rivera, Katrina T. Forest)....Pages 311-320
Blue Native PAGE Analysis of Bacterial Secretion Complexes (Susann Zilkenat, Tobias Dietsche, Julia V. Monjarás Feria, Claudia E. Torres-Vargas, Mehari Tesfazgi Mebrhatu, Samuel Wagner)....Pages 321-351
In Situ Imaging of Bacterial Secretion Systems by Electron Cryotomography (Gregor L. Weiss, João M. Medeiros, Martin Pilhofer)....Pages 353-375
Structural Analysis of Protein Complexes by Cryo Electron Microscopy (Tiago R. D. Costa, Athanasios Ignatiou, Elena V. Orlova)....Pages 377-413
Bacterial Filamentous Appendages Investigated by Solid-State NMR Spectroscopy (Birgit Habenstein, Antoine Loquet)....Pages 415-448
Energy Requirements for Protein Secretion via the Flagellar Type III Secretion System (Marc Erhardt)....Pages 449-457
Identification of Effectors: Precipitation of Supernatant Material (Nicolas Flaugnatti, Laure Journet)....Pages 459-464
Screening for Secretion of the Type VI Secretion System Protein Hcp by Enzyme-Linked Immunosorbent Assay and Colony Blot (Brent S. Weber, Pek Man Ly, Mario F. Feldman)....Pages 465-472
Effector Translocation: Cya Reporter Assay (Suma Chakravarthy, Bethany Huot, Brian H. Kvitko)....Pages 473-487
Monitoring Effector Translocation using the TEM-1 Beta-Lactamase Reporter System (Julie Allombert, Anne Vianney, Xavier Charpentier)....Pages 489-499
Effector Translocation Assay: Differential Solubilization (Irina S. Franco, Sara V. Pais, Nuno Charro, Luís Jaime Mota)....Pages 501-515
Quantitative Determination of Anti-bacterial Activity During Bacterial Co-culture (Juliana Alcoforado Diniz, Birte Hollmann, Sarah J. Coulthurst)....Pages 517-524
Back Matter ....Pages 525-528

✦ Subjects

Bacterial proteins -- Laboratory manuals;Bacterial proteins;Bacteriological Techniques;Bacterial Proteins -- secretion;Clinical Laboratory Techniques