Unit

UNIT 18.4 Native Chemical Ligation of Polypeptides

  1. Julio A. Camarero,
  2. Tom W. Muir

Published Online: 1 MAY 2001

DOI: 10.1002/0471140864.ps1804s15

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Camarero, J. A. and Muir, T. W. 2001. Native Chemical Ligation of Polypeptides. Current Protocols in Protein Science. 15:18.4:18.4.1–18.4.21.

Author Information

  1. The Rockefeller University, New York, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAR 1999

Abstract

The total synthesis and semisynthesis of proteins allows the site-specific incorporation of unnatural amino acids, post-translational modifications, and biophysical/biochemical probes into the target molecule. Among the various chemical and enzymatic approaches available for the synthesis/semisynthesis of proteins, the native chemical ligation technique has proven especially useful and is the exclusive focus of this unit. This unit first discusses how to choose the ligation site(s) in the target protein and then outlines how to obtain the necessary polypeptide building blocks using either chemical synthesis or recombinant DNA expression. Next, the synthesis of a protein by native chemical ligation of two polypeptide fragments is described. The synthesis of a protein from three polypeptide fragments using a sequential native chemical ligation strategy is also described. Support protocols describe how to obtain the necessary polypeptide fragments using either chemical synthesis or recombinant DNA expression.