E-Book

Enzymes as Sensors [electronic resource]

  • Fierke, Carol A. , Thompson, Richard B.
  • Academic Press
  • 2017
Enzymes as Sensors [electronic resource]
  • 자료유형
    단행본
  • 서명/저자사항
    Enzymes as Sensors[electronic resource] / Richard B. Thompson, Carol A. Fierke.
  • 발행사항
    [Cambridge, MA] : Academic Press, 2017.
  • 개인저자
    Fierke, Carol A., Thompson, Richard B.
  • 형태사항
    1 online resource.
  • 총서명
    Methods in Enzymology ; 589
  • ISSN
    0076-6879
  • ISBN
    9780128054062
  • 언어
    영어

목차

Chapter One: Recent Advances in Development of Genetically Encoded Fluorescent Sensors

Abstract
1 Introduction
2 Fluorescent Proteins
3 Sensor Platforms
4 Types of Sensors
5 Conclusion
Chapter Two: Engineering Rugged Field Assays to Detect Hazardous Chemicals Using Spore-Based Bacterial Biosensors

Abstract
1 Introduction
2 Whole Cell-Based Biosensors
3 Design, Creation, and Use of WCBs
4 The Development of Spore-Based Bacterial Biosensors
5 Challenges, Innovations, and Future Directions
6 Conclusions
Acknowledgments
Chapter Three: Engineering BRET-Sensor Proteins

Abstract
1 Introduction
2 LUCIDs and LUMABS: BRET-Sensors for Diagnostic Applications
3 BRET/FRET-Sensors for Intracellular Measurements
4 A Bright Future for BRET-Sensors
Acknowledgments
Chapter Four: Enzymes as Sensors

Abstract
1 Introduction
2 Optical Biosensors
3 Label-Free Detection and Fluorescence-Based Detection
4 Why Choose Biomolecules for Sensing?
5 Enzymes as MREs
6 Thermophilic Enzymes
7 Biological Engineering
8 Conclusions
Acknowledgments
Chapter Five: Integrated Strategies to Gain a Systems-Level View of Dynamic Signaling Networks

Abstract
1 Introduction
2 Strategies to Characterize Global Changes in the Phosphorylation Status of Cellular Proteins
3 Strategies to Track Kinase and Phosphatase Activity Profiles in Living Cells
4 Conclusions
Acknowledgments
Chapter Six: Probing Cdc42 Polarization Dynamics in Budding Yeast Using a Biosensor

Abstract
1 Introduction
2 Generation of a Biosensor of Active Cdc42
3 Detection and Quantification of Cdc42 Activation In Vivo
4 Application of the Cdc42 Biosensor
5 Concluding Remarks
Acknowledgments
Chapter Seven: Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids

Abstract
1 Introduction
2 Site-Specific Incorporation of Unnatural Amino Acids Into Proteins in Live Cells
3 An Unnatural Fluorescent Protein Biosensor for Hydrogen Peroxide
4 Concluding Remarks
Acknowledgments
Chapter Eight: Folding- and Dynamics-Based Electrochemical DNA Sensors

Abstract
1 Introduction
2 “Signal-Off” E-DNA Sensors
3 “Signal-On” E-DNA Sensors
4 Alternate Electrode Substrates
5 Alternate Sensor Fabrication Approaches
6 Conclusion and Outlook
Acknowledgment
Chapter Nine: Construction of Protein-Based Biosensors Using Ligand-Directed Chemistry for Detecting Analyte Binding

Abstract
1 Introduction
2 Expression of GABAAR Consisting of α/β/γ Subunits in HEK293T Cells
3 Design and Synthesis of Labeling Reagents for GABAAR
4 Chemical Labeling of GABAAR Using LDAI Reagent
5 Construction of Fluorescent Biosensors Using Labeled GABAARs
6 Quantitative Analysis of Ligand Affinity Using BFQR-Based Biosensors
7 Summary and Future Directions
Acknowledgments
Chapter Ten: Measuring and Imaging Metal Ions With Fluorescence-Based Biosensors: Speciation, Selectivity, Kinetics, and Other Issues

Abstract
1 Introduction
2 Speciation of Metal Ions of Biological Interest
3 Metal Ion Buffers and Their Formulation
4 Selectivity of Metal Ion Binding to Indicators and Its Assessment
5 Advantages of Biomolecules for Metal Ion Sensing
6 Kinetics and Mechanism of Metal Ion Binding
7 Distribution of Metal Ions Throughout the Cell and Organism
8 Conclusion
Chapter Eleven: Bioapplications of Electrochemical Sensors and Biosensors

Abstract
1 Introduction
2 Electrochemical Sensing in Biological Systems
3 Recent Advancements in Electrode Designs for Biological Monitoring
4 Conclusions and Future Trends
Acknowledgments
Chapter Twelve: A Highly Sensitive Biosensor for ATP Using a Chimeric Firefly Luciferase

Abstract
1 Introduction
2 ATP Detection Protocol
3 Conclusions
Acknowledgments
Chapter Thirteen: Highly Modular Bioluminescent Sensors for Small Molecules and Proteins

Abstract
1 Introduction
2 Sensing Small Molecules
3 Sensing Proteins
4 Toward Application in POC Settings
5 Conclusions
Chapter Fourteen: Sensitive Protein Detection and Quantification in Paper-Based Microfluidics for the Point of Care

Abstract
1 Introduction
2 Paper-Based Devices and Sandwich Lateral Flow Assays
3 Assay Analysis Framework
4 Binding Kinetics in Porous Networks
5 Mechanisms for Protein Binding
6 Optical Methods in Paper
7 Sensitivity Enhancements (LOD)
8 Summary and Next Steps
Chapter Fifteen: Microneedle Enzyme Sensor Arrays for Continuous In Vivo Monitoring

Abstract
1 Introduction
2 Approaches to the Fabrication of Microneedles for Sensing
3 Functionalization of Microneedles for Sensing
4 Methods
Acknowledgments
Chapter Sixteen: Visualization of the Genomic Loci That Are Bound by Specific Multiprotein Complexes by Bimolecular Fluorescence Complementation Analysis on Drosophila Polytene Chromosomes

Abstract
1 Introduction
2 Design of Fusion Proteins for BiFC Analysis
3 Construction and Testing of Plasmid Vectors for BiFC Analysis
4 Production of Drosophila Lines That Express BiFC Fusion Proteins
5 Comparison of the Levels of BiFC Fusion Protein Expression
6 Preparation of Drosophila Larvae for Salivary Gland Isolation
7 Preparation of Polytene Chromosome Spreads
8 Staining and Imaging of Polytene Chromosomes
9 Interpretation of BiFC Complex Binding on Polytene Chromosomes
10 Strategies to Limit the Artifacts Caused by the Differences Between Fusion Proteins and Endogenous Counterparts
11 Effects of BiFC Complex Formation on the Specificity of Chromatin Binding
12 Materials and Solutions