Two-Component Signaling Systems, Part B Methods in Enzymology Series

Multicellular organisms must be able to adapt to cellular events to accommodate prevailing conditions. Sensory-response circuits operate by making use of a phosphorylation control mechanism known as the "two-component system."

Sections in Two-Component Signaling Systems, Part B include:

• Structural Approaches
• Reconstitution of Heterogeneous Systems
• Intracellular Methods and Assays
• Genome-Wide Analyses of Two-Component Systems

Section I: Structural Approaches

[1]: The PICM Chemical Scanning Method for Identifying Domain–Domain and Protein–Protein Interfaces: Applications to the Core Signaling Complex of E. coli Chemotaxis

[2]: Use of Site-Directed Cysteine and Disulfide Chemistry to Probe Protein Structure and Dynamics: Applications to Soluble and Transmembrane Receptors of Bacterial Chemotaxis

[3]: Measuring Distances by Pulsed Dipolar ESR Spectroscopy: Spin-Labeled Histidine Kinases

[4]: Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR

[5]: TonB/TolA Amino-Terminal Domain Modeling

[6]: Functional Dynamics of Response Regulators Using NMR Relaxation Techniques

[7]: The Design and Development of Tar-EnvZ Chimeric Receptors

[8]: Functional and Structural Characterization of EnvZ, an Osmosensing Histidine Kinase of E. coli

[9]: Light Modulation of Histidine-Kinase Activity in Bacterial Phytochromes Monitored by Size Exclusion Chromatography, Crosslinking, and Limited Proteolysis

[10]: A Temperature-Sensing Histidine Kinase—Function, Genetics, and Membrane Topology

[11]: The Regulation of Histidine Sensor Kinase Complexes by Quorum Sensing Signal Molecules

Section II: Reconstitution of Heterogeneous Systems

[12]: Liposome-Mediated Assembly of Receptor Signaling Complexes

[13]: Analyzing Transmembrane Chemoreceptors Using In Vivo Disulfide Formation Between Introduced Cysteines

[14]: Using Nanodiscs to Create Water-Soluble Transmembrane Chemoreceptors Inserted in Lipid Bilayers

[15]: Assays for CheC, FliY, and CheX as Representatives of Response Regulator Phosphatases

[16]: Genetic Dissection of Signaling Through the Rcs Phosphorelay

Section III: Intracellular Methods and Assays

[17]: In Vivo Measurement by FRET of Pathway Activity in Bacterial Chemotaxis

[18]: In Vivo and In Vitro Analysis of the Rhodobacter sphaeroides Chemotaxis Signaling Complexes

[19]: In Vivo Crosslinking Methods for Analyzing the Assembly and Architecture of Chemoreceptor Arrays

[20]: A “Bucket of Light” for Viewing Bacterial Colonies in Soft Agar

[21]: Phenotypic Suppression Methods for Analyzing Intra- and Inter-Molecular Signaling Interactions of Chemoreceptors

[22]: Single-Cell Analysis of Gene Expression by Fluorescence Microscopy

Section IV: Genome-Wide Analyses of Two-Component Systems

[23]: Two-Component Systems of Mycobacterium tuberculosis—Structure-Based Approaches

[24]: Transcriptomic Analysis of ArlRS Two-Component Signaling Regulon, a Global Regulator, in Staphylococcus aureus

[25]: Global Analysis of Two-Component Gene Regulation in H. pylori by Mutation Analysis and Transcriptional Profiling

[26]: Phosphotransfer Profiling: Systematic Mapping of Two-Component Signal Transduction Pathways and Phosphorelays

[27]: Identification of Histidine Phosphorylations in Proteins Using Mass Spectrometry and Affinity-Based Techniques

Subject Index

Author Index

• Presents detailed protocols
• Includes troubleshooting tips