The BAM Complex, 1st ed. 2015 Methods and Protocols Methods in Molecular Biology Series, Vol. 1329

This volume is comprised of a collection of experimental protocols for common techniques and strategies used to study the biogenesis of b-barrel outer membrane proteins in Gram-negative bacteria. The BAM Complex: Methods and Protocols guides readers through methods on the function of the BAM complex, the roles played by each of the individual components, the expression and purification of the components, crystallization and structure determination of the components, and how the individual Bam components may assemble into a functional complex. 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,The BAM Complex: Methods and Protocols will serve as an invaluable reference for those interested in studying the BAM complex.

1. The β-barrel Assembly Machinery Complex

Denisse L. Leyton, Matthew J. Belousoff, and Trevor Lithgow

2. Yeast Mitochondria as a Model System to Study the Biogenesis of Bacterial β-barrel Proteins

Thomas Ulrich, Philipp Oberhettinger, Ingo B. Autenrieth,  and Doron Rapaport

3. Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

Martine P. Bos, Ria Tommassen-van Boxtel, and Jan Tommassen

4. Heat Modifiability of Outer Membrane Proteins from Gram-Negative Bacteria

Nicholas Noinaj,Adam J. Kuszak, and Susan K. Buchanan

5. The Role of a Destabilized Membrane for OMP Insertion

Ashlee M. Plummer, Dennis Gessmann, and Karen G. Fleming

6. Treponema pallidum inGel Microdroplets: A Method for Topological Analysis of BamA (TP0326) and Localization of Rare Outer Membrane Proteins

Amit Luthra, Arvind Anand, and Justin D. Radolf

7. Analyzing the Role of Periplasmic Folding Factors in the Biogenesis of OMPs and Members of the Type V Secretion System

Gustavo Bodelón, Elvira Marín, and Luis Ángel Fernández

8. An In Vitro Assay for Substrate Translocation by FhaC in Liposomes

Enguo Fan, Derrick Norell, and Matthias Müller

9. Measuring Cell-cell Binding Using Flow-Cytometry

Zachary C. Ruhe, Christopher S. Hayes, and David A.

10. Methods to Characterize Folding and Function of BamA Crosslink Mutants

Adam J. Kuszak, Nicholas Noinaj, and Susan K. Buchanan

11. Small Angle X-ray Scattering (SAXS) Characterization of the POTRA Domains of BamA

Pamela Arden Doerner and Marcelo Carlos Sousa

12. Assessing the Outer Membrane Insertion and Folding of Multimeric Transmembrane β-barrel Proteins

Jack C. Leo, Philipp Oberhettinger, and Dirk Linke

13. The Expression, Purification, and Structure Determination of BamA from E. coli

Dongchun Ni and Yihua Huang

14. Expression and Purification of the Individual Bam Components BamB-E

Suraaj Aulakh, Kelly H. Kim, and Mark Paetzel

15. Structure Determination of the BAM Complex Accessory Lipoproteins BamB-E

Kornelius Zeth

16. An In Vitro Assay for Outer Membrane Protein Assembly by the BAM complex

Giselle Roman-Hernandez and Harris D. Bernstein

17. Identification of BamC on the Surface of E. coli

Chaille T. Webb and Trevor Lithgow

18. Construction and Characterization of an E. colibamD Depletion Strain

Dante P. Ricci

19. Expression, Purification and Screening of BamE, a Component of the BAM Complex, for Structural Characterization

Mark Jeeves, Pooja Sridhar, and Timothy J.

20. Purification and Bicelle Crystallization for Structure Determination of the E. coli Outer Membrane Protein TamA

Fabian Gruss, Sebastian Hiller, and Timm Maier

21. Strategies for the Analysis of Bam Recognition Motifs in Outer Membrane Proteins

Nagarajan Paramasivam and Dirk Linke

22. Summary and Future Directions

Nicholas Noinaj and Susan K. Buchanan

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts