Split Inteins

Methods and Protocols

  • Henning D. Mootz

Part of the Methods in Molecular Biology book series (MIMB, volume 1495)

Table of contents

  1. Front Matter
    Pages i-xiv
  2. Eliot L. Osher, Ali Tavassoli
    Pages 27-39
  3. Krishnappa Jagadish, Julio A. Camarero
    Pages 41-55
  4. Shubhendu Palei, Henning D. Mootz
    Pages 77-92
  5. Stefanie Hackl, Alanca Schmid, Christian F. W. Becker
    Pages 93-109
  6. Radhika Borra, Julio A. Camarero
    Pages 111-130
  7. Tobias Schubeis, Madhu Nagaraj, Christiane Ritter
    Pages 147-160
  8. Miguel A. Ramirez, Zhilei Chen
    Pages 161-171
  9. Spencer C. Alford, Connor O’Sullivan, Perry L. Howard
    Pages 197-216
  10. Anam Qudrat, Abdullah Mosabbir, Kevin Truong
    Pages 227-237
  11. Yi-Zong Lee, Wei-Cheng Lo, Shih-Che Sue
    Pages 259-268
  12. Back Matter
    Pages 269-270

About this book

Introduction

This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments.

Keywords

split-intein fusion proteins mutations affinity chromatography elastin-like peptide tags (ELP) peptide macrocycles bioactive cyclotide synthetic intein-extein pieces fluorophores protein trans-splicing toxic enzyme barnase unnatural amino acid mutagenesis

Editors and affiliations

  • Henning D. Mootz
    • 1
  1. 1.Institute of Biochemistry Department of Chemistry and PharmacyUniversity of MuensterMünsterGermany

Bibliographic information

  • DOI https://doi.org/10.1007/978-1-4939-6451-2
  • Copyright Information Springer Science+Business Media New York 2017
  • Publisher Name Humana Press, New York, NY
  • eBook Packages Springer Protocols
  • Print ISBN 978-1-4939-6449-9
  • Online ISBN 978-1-4939-6451-2
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • About this book
Industry Sectors
Chemical Manufacturing
Biotechnology
Consumer Packaged Goods
Pharma