Non-natural amino acids / edited by Tom W. Muir, John N. Abelson. — Amsterdam ; London : Academic, 2009. – (58.17435/C719/v.462)

Contents

    CONTENTS
    
    Contributors
    Preface
    Volumes in Series
    1. Protein Phosphorylation by Semisynthesis: From Paper to Practice
     1. Overview of Protein Phosphorylation
     2. Investigating Protein Phosphorylation with Phosphomimetics
     3. Phosphonate Analogues and Protein Semisynthesis
     4. Methods
     5. Practical Uses of Semisynthetic Phosphoproteins
     6. Future of Protein Semisynthesis in Signaling
     Acknowledgments
     References
    2. Protein Engineering with the Traceless Staudinger Ligation 25
     1. Introduction
     2. Traceless Staudinger Ligation
     3. Choice of Coupling Reagent
     4. Preparation of the Azido Fragment
     5. Preparation of the Phosphinothioester Fragment
     6. Protein Assembly by Orthogonal Chemical Ligations
     7. Prospectus
     Acknowledgments
     References
    3. Replacement of Y730 and Y731 in the α2 Subunit of Escherichia coil Ribonucleotide Reductase with 3-Aminotyrosine using an Evolved Suppressor tRNA/tRNA-Synthetase Pair 45
     1. Introduction
     2. Site-Specific Insertion of Unnatural Amino Acids using the Suppressor tRNA/RS Method
     3. NH2Y， a Y Analogue for Investigating Enzymatic PCET Reactions
     4. Directed Evolution of NH2Y-RS in E. coli
     5. Examination of the Fidelity and Specificity of NH2Y Incorporation in a Protein Expressed in E. coli
     6. Generation of Y730NH2Y-α2 and Y731 NH2Y-α2
     7. Characterization of NH2Y-α2s
     8. Summary
     Acknowledgments
     References
    4. Semisynthesis of Proteins Using Split Inteins
     1. Introduction
     2. Protein Splicing in cis and in trans is Performed by Inteins
     3. Design of Split Inteins and Considerations on the Intein Insertion Site
     4. Materials and Methods
     5. Summary and Conclusion
     Acknowledgments
     References
    5. Expressed Protein Ligation for Metalloprotein Design and Engineering
     1. Introduction
     2. Methods of Selenocysteine Incorporation
     3. Incorporation of Selenocysteine into the Type I Copper Site of Azurin
     4. Tuning the Type I Copper Reduction Potential Using Isostructural Methionine Analogues
     5. Conclusion
     Acknowledgments
     References
    6. Using Expressed Protein Ligation to Probe the Substrate Specificity of Lantibiotic Synthetases
     1. Introduction
     2. Use of Expressed Protein Ligation to Prepare Substrate Analogues
     3. Conclusion
     Acknowledgments
     References
    7. Semisynthesis of K+ Channels
     1. Introduction
     2. Experimental Protocols
     3. Application of Semisynthesis in Investigating the Selectivity Filter of the K+ Channels
     4. Summary
     Acknowledgment
     References
    8. Segmental Isotopic Labeling of Proteins for Nuclear Magnetic Resonance
     1. Introduction
     2. Segmental Isotopic Labeling using Expressed Protein Ligation
     3. Segmental Labeling using Protein Trans-Splicing
     4. Multiple Segment Assembly
     5. Segmental Labeling of C-Terminal SRC Kinase(Csk)
     Acknowledgments
     References
    9. Semisynthesis of Membrane-Attached Prion Proteins
     1. Introduction
     2. Chemical Synthesis of Membrane Anchors
     3. Semisynthesis of rPrPPalm by Expressed Protein Ligation
     4. Semisynthesis of rPrPPalm by Protein Trans-Splicing
     5. Liposome Attachment and Aggregation of rPrPPalm and rPrPGPI
     6. Conclusion
     References
    10. Use of Intein-Mediated Protein Ligation Strategies for the Fabrication of Functional Protein Arrays
     1. Introduction
     2. Intein-Mediated Protein Ligation Strategies
     3. In Vitro， In Vivo and Cell Free Strategies for Protein Biotinylation at the C-Terminal
     4. Methods
     5. Concluding Remarks
     References
    11. Semisynthesis of Ubiquitylated Proteins
     1. Introduction
     2. Semisynthesis of Ubiquitylated Histone H2B
     3. Methods
     4. Synthesis of Photocleavable Ligation Auxiliary
     5. Peptide Synthesis
     6. Generation of Recombinant Protein α-Thioesters
     7. Expressed Protein Ligation
     8. Generation of Ubiquitylated Mononucleosomes
     9. Conclusions
     Acknowledgments
     References
    Author Index
    Subject Index