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Genetically Incorporated Non-Canonical Amino Acids

Methods and Protocols

  • Book
  • © 2023

Overview

Editors:
  1. Yu-Hsuan Tsai

    1. Institute of Molecular Physiology, Shenzhen Bay Laboratory, Shenzhen, China

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  2. Simon J. Elsässer

    1. Laboratory of Synthetic and Systems Biology, Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Solna, Stockholm, Sweden

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2676)

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Genetic Code Expansion in E. coli and Its Applications

    1. Front Matter

      Pages 1-1
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    2. Focused Engineering of Pyrrolysyl-tRNA Synthetase-Based Orthogonal Translation Systems for the Incorporation of Various Noncanonical Amino Acids

      • Nikolaj G. Koch, Nediljko Budisa
      Pages 3-19

    3. Engineering Homogeneous Photoactive Antibody Fragments

      • Thomas Bridge, Amit Sachdeva
      Pages 21-40

    4. Repurposing Photosensitizer Proteins Through Genetic Code Expansion to Facilitate Photo-Biocatalysis

      • Jiangyun Wang, Yan Xia, Xuzhen Guo
      Pages 41-54

    5. Genetic Encoding of a Fluorescent Noncanonical Amino Acid as a FRET Donor for the Analysis of Deubiquitinase Activities

      • Manjia Li, Tao Peng
      Pages 55-67

    6. Creating Selenocysteine-Specific Reporters Using Inteins

      • Christina Z. Chung, Dieter Söll, Natalie Krahn
      Pages 69-86

    7. Protein Expression with Biosynthesized Noncanonical Amino Acids

      • Yong Wang, Wenkang Cai, Boyang Han, Tao Liu
      Pages 87-100

    8. Reprogramming Initiator and Nonsense Codons to Simultaneously Install Three Distinct Noncanonical Amino Acids into Proteins in E. coli

      • Han-Kai Jiang, Jeffery M. Tharp
      Pages 101-116

    9. Encoding Noncanonical Amino Acids into Phage Displayed Proteins

      • Cristina Díaz-Perlas, Montserrat Escobar-Rosales, Charles W. Morgan, Benjamí Oller-Salvia
      Pages 117-129

    10. Genetically Encoded Noncanonical Amino Acids in Proteins to Investigate Lysine Benzoylation

      • An-Di Guo, Xiao-Hua Chen
      Pages 131-146

    11. Semisynthesis of Glutamine-Methylated Proteins Enabled by Genetic Code Expansion

      • Weimin Xuan, Xiaochen Yang
      Pages 147-156

  3. Applications in Mammalian Cells

    1. Front Matter

      Pages 157-157
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    2. Genetic Code Expansion in Mammalian Cells

      • Zhigang Wu, Jie Wang
      Pages 159-167

    3. Generation of Amber Suppression Cell Lines Using CRISPR-Cas9

      • Birthe Meineke, Simon J. Elsässer
      Pages 169-180

    4. Genetic Code Expansion in Mammalian Cells Through Quadruplet Codon Decoding

      • Yan Chen, Tianyu Gao, Xinyuan He, Wei Niu, Jiantao Guo
      Pages 181-190

    5. Genetically Encoded 1,2-Aminothiol for Site-Specific Modification of a Cellular Membrane Protein via TAMM Condensation

      • Han Sun, Yang Huang, Yu-Hsuan Tsai
      Pages 191-199

    6. Conformational GPCR BRET Sensors Based on Bioorthogonal Labeling of Noncanonical Amino Acids

      • Maria Kowalski-Jahn, Hannes Schihada, Gunnar Schulte
      Pages 201-213

    7. Selective Inhibition of Kinase Activity in Mammalian Cells by Bioorthogonal Ligand Tethering

      • Jinghao Chen, Yang Huang, Wen-Biao Gan, Yu-Hsuan Tsai
      Pages 215-232

    8. Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells

      • Xinyuan He, Yan Chen, Jiantao Guo, Wei Niu
      Pages 233-243
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Keywords

About this book

This detailed volume explores non-canonical amino acids (ncAAs) through their site-specific incorporation by genetic code expansion (GCE). The collection provides a broad resource of methods for implementing GCE in E. coli, mammalian cells, and animals, highlighting specific applications ranging from fluorescence labeling to photocontrol and the study of protein post-translational modification. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Genetically Incorporated Non-Canonical Amino Acids: Methods and Protocols serves as an ideal source of methodologies that can be adapted and extended, migrated to different model systems, and combined in new ways to help explore a wide range of biological questions and to augment industrial and pharmaceutical protein engineering.

Editors and Affiliations

  • Institute of Molecular Physiology, Shenzhen Bay Laboratory, Shenzhen, China

    Yu-Hsuan Tsai

  • Laboratory of Synthetic and Systems Biology, Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Solna, Stockholm, Sweden

    Simon J. Elsässer

Bibliographic Information

  • Book Title: Genetically Incorporated Non-Canonical Amino Acids

  • Book Subtitle: Methods and Protocols

  • Editors: Yu-Hsuan Tsai, Simon J. Elsässer

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-3251-2

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2023

  • Hardcover ISBN: 978-1-0716-3250-5Published: 06 June 2023

  • Softcover ISBN: 978-1-0716-3253-6Due: 07 July 2023

  • eBook ISBN: 978-1-0716-3251-2Published: 05 June 2023

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 286

  • Number of Illustrations: 1 b/w illustrations

  • Topics: Chemistry/Food Science, general, Biochemistry, general

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