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Synthetic Protein Switches

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Cover of 'Synthetic Protein Switches'

Table of Contents

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    Book Overview
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    Chapter 1 Synthetic Protein Switches: Theoretical and Experimental Considerations
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    Chapter 2 Construction of Allosteric Protein Switches by Alternate Frame Folding and Intermolecular Fragment Exchange
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    Chapter 3 Construction of Protein Switches by Domain Insertion and Directed Evolution
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    Chapter 4 Catalytic Amyloid Fibrils That Bind Copper to Activate Oxygen
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    Chapter 5 Ancestral Protein Reconstruction and Circular Permutation for Improving the Stability and Dynamic Range of FRET Sensors
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    Chapter 6 Method for Developing Optical Sensors Using a Synthetic Dye-Fluorescent Protein FRET Pair and Computational Modeling and Assessment
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    Chapter 7 Rational Design and Applications of Semisynthetic Modular Biosensors: SNIFITs and LUCIDs
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    Chapter 8 Ultrasensitive Firefly Luminescent Intermediate-Based Protein-Protein Interaction Assay (FlimPIA) Based on the Functional Complementation of Mutant Firefly Luciferases
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    Chapter 9 Quantitative and Dynamic Imaging of ATM Kinase Activity
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    Chapter 10 Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains
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    Chapter 11 Protein and Protease Sensing by Allosteric Derepression
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    Chapter 12 DNA-Specific Biosensors Based on Intramolecular β-Lactamase-Inhibitor Complex Formation
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    Chapter 13 Engineering and Characterizing Synthetic Protease Sensors and Switches
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    Chapter 14 Characterizing Dynamic Protein–Protein Interactions Using the Genetically Encoded Split Biosensor Assay Technique Split TEV
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    Chapter 15 Development of a Synthetic Switch to Control Protein Stability in Eukaryotic Cells with Light
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    Chapter 16 Light-Regulated Protein Kinases Based on the CRY2-CIB1 System
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    Chapter 17 Yeast-Based Screening System for the Selection of Functional Light-Driven K+ Channels
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    Chapter 18 Primer-Aided Truncation for the Creation of Hybrid Proteins
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    Chapter 19 Engineering Small Molecule Responsive Split Protein Kinases
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    Chapter 20 Directed Evolution Methods to Rewire Signaling Networks
Attention for Chapter 6: Method for Developing Optical Sensors Using a Synthetic Dye-Fluorescent Protein FRET Pair and Computational Modeling and Assessment
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Chapter title
Method for Developing Optical Sensors Using a Synthetic Dye-Fluorescent Protein FRET Pair and Computational Modeling and Assessment
Chapter number 6
Book title
Synthetic Protein Switches
Published in
Methods in molecular biology, March 2017
DOI 10.1007/978-1-4939-6940-1_6
Pubmed ID
Book ISBNs
978-1-4939-6938-8, 978-1-4939-6940-1
Authors

Joshua A. Mitchell, William H. Zhang, Michel K. Herde, Christian Henneberger, Harald Janovjak, Megan L. O’Mara, Colin J. Jackson

Editors

Viktor Stein

Abstract

Biosensors that exploit Förster resonance energy transfer (FRET) can be used to visualize biological and physiological processes and are capable of providing detailed information in both spatial and temporal dimensions. In a FRET-based biosensor, substrate binding is associated with a change in the relative positions of two fluorophores, leading to a change in FRET efficiency that may be observed in the fluorescence spectrum. As a result, their design requires a ligand-binding protein that exhibits a conformational change upon binding. However, not all ligand-binding proteins produce responsive sensors upon conjugation to fluorescent proteins or dyes, and identifying the optimum locations for the fluorophores often involves labor-intensive iterative design or high-throughput screening. Combining the genetic fusion of a fluorescent protein to the ligand-binding protein with site-specific covalent attachment of a fluorescent dye can allow fine control over the positions of the two fluorophores, allowing the construction of very sensitive sensors. This relies upon the accurate prediction of the locations of the two fluorophores in bound and unbound states. In this chapter, we describe a method for computational identification of dye-attachment sites that allows the use of cysteine modification to attach synthetic dyes that can be paired with a fluorescent protein for the purposes of creating FRET sensors.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 8 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 8 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 2 25%
Student > Ph. D. Student 1 13%
Unspecified 1 13%
Student > Bachelor 1 13%
Professor > Associate Professor 1 13%
Other 0 0%
Unknown 2 25%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 2 25%
Unspecified 1 13%
Agricultural and Biological Sciences 1 13%
Chemistry 1 13%
Neuroscience 1 13%
Other 0 0%
Unknown 2 25%