Enzymes as Sensors - 1st Edition - ISBN: 9780128054062, 9780128092910

Enzymes as Sensors, Volume 589

1st Edition

Serial Volume Editors: Richard Thompson Carol A. Fierke
eBook ISBN: 9780128092910
Hardcover ISBN: 9780128054062
Imprint: Academic Press
Published Date: 24th March 2017
Page Count: 530
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Table of Contents

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

Description

Enzymes as Sensors, Volume 589, the latest release in the Methods in Enzymology series, covers a variety of topics, including advances in genetically coded fluorescent sensors, enzymes as sensors, and bioapplications of electrochemical sensors and biosensors. Users will find a comprehensive discussion of timely topics that presents a micro-level delivery of specific content related to the study of enzymes in sensors. New to this edition are highly specialized chapters on integrated strategies for gaining a systems level view of dynamic signaling networks, sensitive protein detection and quantification in paper-based microfluidics for point-of-care, and microneedle enzyme sensor arrays for continuous in vivo monitoring.

This state-of-the-art series is ideal for those interested in the latest information on enzymology, with this edition focusing on sensors and their role in enzymes.

Key Features

  • Contain contributions from leading authorities
  • Informs and updates on all the latest developments in the field

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists


Details

No. of pages:
530
Copyright:
© Academic Press 2017
Published:
Imprint:
Academic Press
eBook ISBN:
9780128092910
Hardcover ISBN:
9780128054062

About the Serial Volume Editors

Richard Thompson Serial Volume Editor

Richard B. Thompson is Associate Professor at Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, U.S.A.

Affiliations and Expertise

Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, USA

Carol A. Fierke Serial Volume Editor

Carol Fierke is Professor and Chair at the Department of Chemistry and Professor of Biological Chemistry at the Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, U.S.A.

Affiliations and Expertise

Department of Chemistry and Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, USA