Bacterial Multidrug Exporters

This detailed volume utilizes our current understanding of the structural basis of multidrug recognition and multidrug efflux mechanisms to provide protocols involving these vital intrinsic membrane proteins widely distributed in bacteria. Beginning with protocols for the structural analysis of bacterial multidrug exporters, the book continues with sections on biochemical and bioengineering analysis, computational analysis, biomedical approaches, as well as advanced technologies expected to be applied to multidrug efflux transport studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Bacterial Multidrug Exporters: Methods and Protocols serves as an ideal guide to this fastmoving field of study.

Includes cutting-edge methods and protocols for the study of bacterial multidrug exporters Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt
High Resolution Crystallographic Analysis of AcrB Using Designed Ankyrin Repeat Proteins (DARPins).- Crystallographic Analysis of Drug and Inhibitor-Binding Structure of RND-Type Multidrug Exporter AcrB in Physiologically-Relevant Asymmetric Crystals.- Crystallographic Analysis of MATE-Type Multidrug Exporter with Its Inhibitors.- Crystallographic Analysis of the CusBA Heavy-Metal Efflux Complex of Escherichia coli.- Purification of AcrAB-TolC Multidrug Efflux Pump for Cryo-EM Analysis.- NMR Spectroscopy Approach to Study the Structure, Orientation, and Mechanism of the Multidrug Exporter EmrE.- Generation of Conformation-Specific Antibody Fragments for Crystallization of the Multidrug Resistance Transporter MdfA.- Biochemical Reconstitution and Characterization of Multi-Component Drug Efflux Transporters.- Covalently Linked Trimers of RND (Resistance-Nodulation-Division) Efflux Transporters to Study Their Mechanism of Action: Escherichia coli AcrB Multidrug Exporter as an Example.- Determining Ligand Path through a Major Drug Transporter, AcrB, in Escherichia coli.- Molecular Modeling of Multi-Drug Properties of Resistance Nodulation Division (RND) Transporters.- A Transcriptomic Approach to Identify Novel Drug Efflux Pumps in Bacteria.- Regulation of the Expression of Bacterial Multidrug Exporters by Two-Component Signal Transduction Systems.- Study of the Expression of Bacterial Multidrug Efflux Pumps in Anaerobic Conditions.- Identification of a Staphylococcus aureusEfflux Pump Regulator Using a DNA-Protein Affinity Technique.- High Throughput Flow Cytometry Screening of Multidrug Efflux Systems.- Single-Molecule Analysis of Membrane Transporter Activity by Means of a Microsystem.- Large Scale Femtoliter Droplet Array for Single Cell Efflux Assay of Bacteria.- Reconstitution and Transport Analysis of Eukaryotic Transporters in the Post-Genomic Era.<p