UNIT 28.8 Chemical Methods and Approaches to the Regioselective Formation of Multiple Disulfide Bonds

  1. Shigeru Shimamoto1,
  2. Hidekazu Katayama2,
  3. Masaki Okumura3,
  4. Yuji Hidaka1

Published Online: 1 APR 2014

DOI: 10.1002/0471140864.ps2808s76

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Shimamoto, S., Katayama, H., Okumura, M. and Hidaka, Y. 2014. Chemical Methods and Approaches to the Regioselective Formation of Multiple Disulfide Bonds. Current Protocols in Protein Science. 76:28.8:28.8.1–28.8.28.

Author Information

  1. 1

    Faculty of Science and Engineering, Kinki University, Osaka, Japan

  2. 2

    Department of Applied Biochemistry, School of Engineering, Tokai University, Kanagawa, Japan

  3. 3

    Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Miyagi, Japan

Publication History

  1. Published Online: 1 APR 2014


Disulfide-bond formation plays an important role in the stabilization of the native conformation of peptides and proteins. In the case of multidisulfide-containing peptides and proteins, numerous folding intermediates are produced, including molecules that contain non-native and native disulfide bonds during in vitro folding. These intermediates can frequently be trapped covalently during folding and subsequently analyzed. The structural characterization of these kinetically trapped disulfide intermediates provides a clue to understanding the oxidative folding pathway. To investigate the folding of disulfide-containing peptides and proteins, in this unit, chemical methods are described for regulating regioselective disulfide formation (1) by using a combination of several types of thiol protecting groups, (2) by incorporating unique SeCys residues into a protein or peptide molecule, and (3) by combining with post-translational modification. Curr. Protoc. Protein Sci. 76:28.8.1-28.8.28. © 2014 by John Wiley & Sons, Inc.


  • folding;
  • intermediate;
  • disulfide;
  • diselenide;
  • cysteine;
  • selenocysteine;
  • post-translation;
  • native chemical ligation;
  • topological isomer