This second edition volume expands on the previous edition with updates on the latest advancements in the field that span from atomic-resolution structural analysis to whole-organism functional studies. The chapters in this book are organized into four parts. Part One: Structural Analysis Methods presents cutting-edge approaches for determining high-resolution structures of multidrug exporters, and covers the full spectrum of structural biology techniques, from solution NMR spectroscopy for studying PfMATE to single-particle cryo-EM analysis of TolC. Part Two: Biochemical Analysis and Reconstitution focuses on functional characterization of multidrug exports and looks at methods such as site-specific fluorescent labeling of AcrB and analytical ultracentrifugation for studying oligomeric states. Part Three: Computational and Simulation Approaches showcases the power of in silico methods in multidrug exporter research such as molecular dynamics simulations of conformational changes in MdfA. Part Four: Genetic Tools and Functional Studies presents a diverse array of methods for investigating multidrug exporters in their biological context. This section explores high-throughput gene mapping using TraDISort, evaluation of efflux pump inhibitors through growth curve analysis, and cloning and characterization protocols. 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.Comprehensive and cutting-edge, Bacterial Multidrug Exporters: Methods and Protocols, Second Edition is a valuable resource for novel researchers entering this field and established investigators seeking to expand their methodological range.

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 tothis fastmoving field of study.

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.

This second edition volume expands on the previous edition with updates on the latest advancements in the field that span from atomic-resolution structural analysis to whole-organism functional studies. The chapters in this book are organized into four parts. Part One: Structural Analysis Methods presents cutting-edge approaches for determining high-resolution structures of multidrug exporters, and covers the full spectrum of structural biology techniques, from solution NMR spectroscopy for studying PfMATE to single-particle cryo-EM analysis of TolC. Part Two: Biochemical Analysis and Reconstitution focuses on functional characterization of multidrug exports and looks at methods such as site-specific fluorescent labeling of AcrB and analytical ultracentrifugation for studying oligomeric states. Part Three: Computational and Simulation Approaches showcases the power of in silico methods in multidrug exporter research such as molecular dynamics simulations of conformational changes in MdfA. Part Four: Genetic Tools and Functional Studies presents a diverse array of methods for investigating multidrug exporters in their biological context. This section explores high-throughput gene mapping using TraDISort, evaluation of efflux pump inhibitors through growth curve analysis, and cloning and characterization protocols. 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.Comprehensive and cutting-edge, Bacterial Multidrug Exporters: Methods and Protocols, Second Edition is a valuable resource for novel researchers entering this field and established investigators seeking to expand their methodological range.

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 tothis fastmoving field of study.

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.