US 6,184,344 B2 · Synthesis of proteins by native chemical ligation

Abstract

Proteins of moderate size having native peptide backbones are produced by a method of native chemical ligation. Native chemical ligation employs a chemoselective reaction of two unprotected peptide segments to produce a transient thioester-linked intermediate. The transient thioester-linked intermediate then spontaneously undergoes a rearrangement to provide the full length ligation product having a native peptide bond at the ligation site. Full length ligation products are chemically identical to proteins produced by cell free synthesis. Full length ligation products may be refolded and/or oxidized, as allowed, to form native disulfide-containing protein molecules. The technique of native chemical ligation is employable for chemically synthesizing full length proteins.

Claims7

Dependent claims

A method for ligating a first oligopeptide with a second oligopeptide end to end for producing an oligopeptide product, the method comprising the following steps:

Step A:
admixing the first and second oligopeptides in a reaction solution including a catalytic thiol, the first oligopeptide including a C-terminal thioester, the second oligopeptide including an N-terminal cysteine having an unoxidized sulfhydryl side chain;
then
Step B:
condensing the unoxidized sulfhydryl side chain of the N-terminal cysteine with the C-terminal thioester for producing an intermediate oligopeptide linking the first and second oligopeptides with a β-aminothioester bond;
and then
Step C:
rearranging the β-aminothioester bond of the intermedite oligopeptide of said Step B for producing the oligopeptide product linking the first and second oligopeptides with an amide bond.

5 more elements

Dependencies are inferred from the claim text.

Synthesis of proteins by native chemical ligation

Life of this patent

Synthesis of proteins by native chemical ligation

Life of this patent

Abstract

Claims7