Description / Table of Contents:
Synthetic Protein Switches: Theoretical and Experimental Considerations -- Construction of Allosteric Protein Switches by Alternate Frame Folding and Intermolecular Fragment Exchange -- Construction of Protein Switches by Domain Insertion and Directed Evolution -- Catalytic Amyloid Fibrils that Bind Copper to Activate Oxygen -- Ancestral Protein Reconstruction and Circular Permutation for Improving the Stability and Dynamic Range of FRET Sensors -- Method for Developing Optical Sensors Using a Synthetic Dye-Fluorescent Protein FRET Pair and Computational Modelling and Assessment -- Rational Design and Applications of Semi-Synthetic Modular Biosensors: SNIFITs and LUCIDs -- Ultrasensitive Firefly Luminescent Intermediate-Based Protein-Protein Interaction Assay (FlimPIA) Based on the Functional Complementation of Mutant Firefly Luciferases -- Quantitative and Dynamic Imaging of ATM Kinase Activity -- Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains -- Protein and Protease Sensing by Allosteric Derepression -- DNA-Specific Biosensors Based on Intramolecular β-Lactamase-Inhibitor Complex Formation -- Engineering and Characterizing Synthetic Protease Sensors and Switches -- Characterizing Dynamic Protein-Protein Interactions Using the Genetically Encoded Split Biosensor Assay Technique Split TEV -- Development of a Synthetic Switch to Control Protein Stability in Eukaryotic Cells with Light -- Light-Regulated Protein Kinases Based on the CRY2-CIB1 System -- Yeast-Based Screening System for the Selection of Functional Light-Driven K+ Channels -- Primer-Aided Truncation for the Creation of Hybrid Proteins -- Engineering Small Molecule Responsive Split-Protein Kinases -- Directed Evolution Methods to Rewire Signaling Networks
This detailed book provides state-of-the-art protocols to facilitate the construction of synthetic protein switches for a variety of applications in biotechnology and basic research. Beginning with general strategies, the volume continues with coverage of peptide switches, fluorescent and bioluminescent sensors, β-lactamase sensors, proteolytic sensors, optogenetic switches, as well as cellular signaling switches. Written for the highly successful Methods in Molecular Biology series, 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 practical, Synthetic Protein Switches: Methods and Protocols serves as a useful reference for life scientists from diverse research fields that range from traditional, discovery-centered disciplines, like cancer research, to newly emerging disciplines, such as synthetic biology
XI, 341 p. 59 illus., 31 illus. in color. : online resource.