UID:
almahu_9947366525102882
Format:
1 online resource (646 p.)
ISBN:
1-281-02881-9
,
9786611028817
,
0-08-054946-2
Series Statement:
Methods in enzymology, volume 423
Content:
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 include:Structural ApproachesReconstitution of Heterogeneous SystemsIntracellular Methods and AssaysGenome-Wide Analyses of Two-Component SystemsPresents detailed protocols Includes troubleshooting tips
Note:
Description based upon print version of record.
,
Cover; Copyright Page; Table of Contents; Contributors to Volume 423; Volume in Series; Section I: Structural Approaches; Chapter 1: The PICM Chemical Scanning Method for Identifying Domain-Domain and Protein-Protein Interfaces: Applications to the Core Signaling Complex of E.Coli Chemotaxis; Introduction; Comparison of the PICM Method with Other Scanning Approaches; PICM Studies of the Core Signaling Complex of Bacterial Chemotaxis; Generalizing the PICM Method to Map Docking Sites in Other Systems; Incorporation of an Affinity Tag and Creation of a Cysless Protein
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Choice of Positions for Cys Incorporation and Creation of a Mutant LibrarySelection of a Cys-Specific Probe for Chemical Modification; Probe Labeling and Purification of the Single Cys Mutants; Quantitation of Probe Coupling; Measuring Functional Effects of Cys Substitution and Bulky Probe Coupling; Interpretation of Results-Mapping Out Docking Sites; Acknowledgments; References; Chapter 2: Use of Site-Directed Cysteine and Disulfide Chemistry to Probe Protein Structure and Dynamics: Applications to Soluble and Transmembrane Receptors of Bacterial Chemotaxis; Introduction
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Site-Directed Cysteine and Disulfide Chemistry: HistorySite-Directed Cysteine and Disulfide Chemistry: Applications and Limitations; Incorporation of an Affinity Tag and Creation of a Cysless Protein; Choice of Positions for Cys Incorporation and Creation of a Mutant Library; Analysis of 2degStructure by Chemical Reactivity Scanning; Disulfide Mapping of Spatial Proximity and Conformational Changes; Disulfide Trapping of Thermal Backbone and Domain Motions; Acknowledgments; References; Chapter 3: Measuring Distances by Pulsed Dipolar ESR Spectroscopy: Spin-Labeled Histidine Kinases
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IntroductionDipolar ESR Spectroscopy; Case Study: PDS Reconstruction of Histidine Kinases Signaling Complex; Concluding Remarks; Acknowledgments; References; Chapter 4: Rigid Body Refinement of Protein Complexes with Long-Range Distance Restraints from Pulsed Dipolar ESR; Introduction; Method; Initial Conformation of the Complex; Results; Discussion; Acknowledgments; References; Chapter 5: TonB/TolA Amino-Terminal Domain Modeling; Introduction; Alanyl Replacement; TonB/TolA Chimeras; Acknowledgments; References
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Chapter 6: Functional Dynamics of Response Regulators Using NMR Relaxation TechniquesIntroduction; The Experimental Setup; Two-State Allosteric Activation Identified by NMR Chemical Shift Analysis; Two-State Allosteric Activation Buttressed by Standard NMR Relaxation Experiments; A New Approach for Quantitative Analysis of Microsecond Protein Dynamics; Conclusions; Acknowledgments; References; Chapter 7: The Design and Development of Tar-EnvZ Chimeric Receptors; Introduction; Construction of Tar-EnvZ Chimeric Protein, Taz; Asp-Dependent Induction of ompC-lacZ Fusion Gene by Taz and OmpR
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Phenotype Analysis of the Taz Construct
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English
Additional Edition:
ISBN 0-12-373852-0
Language:
English
