Linkers in Biomacromolecules
- 1st Edition, Volume 647 - January 20, 2021
- Imprint: Academic Press
- Editor: Maarten Merkx
- Language: English
- Hardback ISBN:9 7 8 - 0 - 1 2 - 8 2 0 8 1 8 - 2
- eBook ISBN:9 7 8 - 0 - 1 2 - 8 2 0 8 2 3 - 6
Linkers in Biomacromolecules, Volume 647 in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Ne… Read more
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Request a sales quoteLinkers in Biomacromolecules, Volume 647 in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this updated release include chapters devoted to ser-gly linkers + models, sergGly- EAAAK linkers, Effect of crowding on linker behavior, Protathether. Experimental approach to quickly test different linkers in fusion proteins, Linkers in enzyme fusions/cascade catalysis, Linker dependence of avidity in multivalent interactions between disordered proteins, ER/K α-helix linker, The effect of chirality and steric hindrance on intrinsic backbone conformational propensities: tools for protein design, Linker design in protein-based nanostructures, and much more. Linkers in Biomacromolecules, Volume 647 in the Methods in Enzymology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this updated release include chapters devoted to ser-gly linkers + models, sergGly- EAAAK linkers, Effect of crowding on linker behavior, Protathether. Experimental approach to quickly test different linkers in fusion proteins, Linkers in enzyme fusions/cascade catalysis, Linker dependence of avidity in multivalent interactions between disordered proteins, ER/K α-helix linker, The effect of chirality and steric hindrance on intrinsic backbone conformational propensities: tools for protein design, Linker design in protein-based nanostructures, and much more.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in the Methods in Enzymology series
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover image
- Title page
- Table of Contents
- Copyright
- Contributors
- Preface
- Chapter One: Understanding and applications of Ser/Gly linkers in protein engineering
- Abstract
- 1: Introduction
- 2: Theoretical description of Ser/Gly linkers using random coil models
- 3: Quantitative understanding of Ser/Gly linker behavior
- 4: Understanding and tuning binding properties of multidomain proteins containing Ser/Gly linkers
- 5: Summary and conclusions
- Acknowledgments
- Chapter Two: Design and construction of chimeric linker library with controllable flexibilities for precision protein engineering
- Abstract
- 1: Introduction
- 2: Engineering of protein linkers
- 3: Design and construction of chimeric linker library
- 4: Measuring the physical length of chimeric linkers with FRET
- 5: Application of chimeric linker library in fusion proteins
- 6: Summary
- Acknowledgments
- Chapter Three: Engineering crowding sensitivity into protein linkers
- Abstract
- 1: Introduction
- 2: The dependence of crowding effects on linker size
- 3: The dependence of crowding effects on linker flexibility
- 4: How do interactions within the linker influence the effect of crowding?
- 5: How do associative interactions of the linker with sticky crowders modulate the steric crowding effect?
- 6: Other crowding effects on linkers
- 7: Protocol
- 8: Maximizing linker compressibility by crowding in cells
- 9: Preventing crowding sensitivity
- 10: Methods in buffer
- 11: Methods in E. coli
- Acknowledgments
- Chapter Four: protaTETHER: A method for the incorporation of linkers in biomacromolecules
- Abstract
- 1: Introduction
- 2: Applications of linkers in biomacromolecules and protein fusions
- 3: Generation and incorporation of linkers with protaTETHER
- 4: Protocol
- 5: Quantification of protaTETHER results
- 6: Summary
- Acknowledgments
- Chapter Five: Optimizing the linker length for fusing an alcohol dehydrogenase with a cyclohexanone monooxygenase
- Abstract
- 1: Introduction
- 2: General method and statistical analysis
- 3: Molecular design of linker variants
- 4: Purification of Hisx6 tagged proteins
- 5: UV–visible spectral analysis of TmCHMO
- 6: Apparent melting temperature determination
- 7: Evaluation of the linker size on the enzyme activity
- 8: Evaluation of fused variants in small-scale bioconversions
- 9: Summary and conclusions
- Acknowledgments
- Chapter Six: Predicting the effect of disordered linkers on effective concentrations and avidity with the “Ceff calculator” app
- Abstract
- 1: Introduction
- 2: Defining linker length and binding site spacing
- 3: Motivation for simplified linker models
- 4: Modeling intramolecular and bivalent binding
- 5: Limitations of WLC models
- 6: Summary
- Acknowledgments
- Chapter Seven: ER/K-link—Leveraging a native protein linker to probe dynamic cellular interactions
- Abstract
- 1: Structure, stability and functional characteristics of ER/K linkers
- 2: ER/K-linked sensors—Mechanism, molecular cloning, protein expression, and measurement of interaction strengths
- 3: Biological insights using ER/K-linked sensors
- 4: Future perspectives
- Acknowledgments
- Chapter Eight: Design of helical linkers for fusion proteins and protein-based nanostructures
- Abstract
- 1: Introduction
- 2: Design of helical linkers for fusion proteins
- 3: Design of linkers for protein-based nanostructures
- 4: Methods
- 5: Concluding remarks
- Acknowledgments
- Chapter Nine: Synthetic protein switches: Combinatorial linker engineering with iFLinkC
- Abstract
- 1: Introduction
- 2: Combinatorial strategies for linker engineering
- 3: Combinatorial linker engineering with iFLinkC
- 4: Protocol
- 5: Summary
- Acknowledgments
- Chapter Ten: Enzyme-DNA chimeras: Construction, allostery, applications
- Abstract
- 1: Introduction
- 2: Enzyme-DNA chimera
- 3: Principles of designing enzyme-DNA chimeras
- 4: Design and synthesize enzyme-DNA chimeras
- 5: Discussion
- 6: Summary
- Acknowledgments
- Chapter Eleven: The Spy that links: Creation of nonlinear protein architectures and materials using SpyTag/SpyCatcher chemistry
- Abstract
- 1: Introduction
- 2: Construct design
- 3: Materials and equipment
- 4: Methods
- 5: Summary
- Edition: 1
- Volume: 647
- Published: January 20, 2021
- Imprint: Academic Press
- No. of pages: 318
- Language: English
- Hardback ISBN: 9780128208182
- eBook ISBN: 9780128208236
MM
Maarten Merkx
Maarten Merkx is at Department of Biomedical Engineering, Eindhoven University of Technology, The Netherlands
Affiliations and expertise
Department of Biomedical Engineering, Eindhoven University of Technology, The NetherlandsRead Linkers in Biomacromolecules on ScienceDirect