Synthetic Protein Switches

1. Front Matter

   Pages i-xi

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2. General Strategies and Considerations

   1. Front Matter

      Pages 1-1

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   2. Synthetic Protein Switches: Theoretical and Experimental Considerations

      • Viktor Stein

      Pages 3-25

   3. Construction of Allosteric Protein Switches by Alternate Frame Folding and Intermolecular Fragment Exchange

      • Jeung-Hoi Ha, Stewart N. Loh

      Pages 27-41

   4. Construction of Protein Switches by Domain Insertion and Directed Evolution

      • Lucas F. Ribeiro, Tiana D. Warren, Marc Ostermeier

      Pages 43-55

3. Peptide Switches

   1. Front Matter

      Pages 57-57

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   2. Catalytic Amyloid Fibrils That Bind Copper to Activate Oxygen

      • Alex Sternisha, Olga Makhlynets

      Pages 59-68

4. Fluorescent and Bioluminescent Sensors

   1. Front Matter

      Pages 69-69

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   2. Ancestral Protein Reconstruction and Circular Permutation for Improving the Stability and Dynamic Range of FRET Sensors

      • Ben E. Clifton, Jason H. Whitfield, Inmaculada Sanchez-Romero, Michel K. Herde, Christian Henneberger, Harald Janovjak et al.

      Pages 71-87

   3. Method for Developing Optical Sensors Using a Synthetic Dye-Fluorescent Protein FRET Pair and Computational Modeling and Assessment

      • Joshua A. Mitchell, William H. Zhang, Michel K. Herde, Christian Henneberger, Harald Janovjak, Megan L. O’Mara et al.

      Pages 89-99

   4. Rational Design and Applications of Semisynthetic Modular Biosensors: SNIFITs and LUCIDs

      • Helen Farrants, Julien Hiblot, Rudolf Griss, Kai Johnsson

      Pages 101-117

   5. Ultrasensitive Firefly Luminescent Intermediate-Based Protein-Protein Interaction Assay (FlimPIA) Based on the Functional Complementation of Mutant Firefly Luciferases

      • Yuki Ohmuro-Matsuyama, Hiroshi Ueda

      Pages 119-130

   6. Quantitative and Dynamic Imaging of ATM Kinase Activity

      • Shyam Nyati, Grant Young, Brian Dale Ross, Alnawaz Rehemtulla

      Pages 131-145

5. β-lactamase Sensors

   1. Front Matter

      Pages 147-147

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   2. Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains

      • Hiroto Iwai, Miki Kojima-Misaizu, Jinhua Dong, Hiroshi Ueda

      Pages 149-165

   3. Protein and Protease Sensing by Allosteric Derepression

      • Hui Chin Goh, Farid J. Ghadessy, Saurabh Nirantar

      Pages 167-177

   4. DNA-Specific Biosensors Based on Intramolecular β-Lactamase-Inhibitor Complex Formation

      • Wouter Engelen, Maarten Merkx

      Pages 179-194

6. Proteolytic Sensors

   1. Front Matter

      Pages 195-195

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   2. Engineering and Characterizing Synthetic Protease Sensors and Switches

      • Viktor Stein, Kirill Alexandrov

      Pages 197-218

   3. Characterizing Dynamic Protein–Protein Interactions Using the Genetically Encoded Split Biosensor Assay Technique Split TEV

      • Jan P. Wintgens, Moritz J. Rossner, Michael C. Wehr

      Pages 219-238

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.

• Bioluminescent sensors
• Fluorescent sensors
• Synthetic biology
• Next-generation therapeutics
• Peptide switches
• β-lactamase
• Optogenetic switches
• Biomolecular analytes

• Fachbereich Biologie, Technische Universität Darmstadt, Darmstadt, Germany

  Viktor Stein

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