Patent ID: 6307018
Filing Date: 2001-10-23
Classification: C07K

Abstract:
A method for ligating a first oligopeptide with a second oligopeptide end-to-end to produce an oligopeptide product, the method comprising the following steps:Step A: admixing the first and second oligopeptides in a reaction solution including a thiol that is present in a catalytic amount, the first oligopeptide including a C-terminal residue having a C-termninal thioester, the second oligopeptide including an N-terminal amino acid residue having a secondary amino group, the secondary amino group being attached to an N-linked auxiliary functional group having an unoxidized sulfhydryl moiety, the C-terminal residue being glycine when the N- terminal residue is non-glycine, the N-terminal residue being glycine when the C-terminal residue is non-glycine;Step B: condensing the unoxidized sulfhydryl moiety of the N-linked auxiliary functional group on the first oligopeptide with the C-terminal thioester of the second oligopeptide for a time and under conditions effective to produce an intermediate oligopeptide which spontaneously rearranges to form the oligopeptide product,wherein said intermediate oligopeptide consists of the first oligopeptide and the second oligopeptide bonded together via an aminothioester bond, and said oligopeptide product consists of the first oligopeptide and the second oligopeptide bonded together via an amide bond, said amide bond bearing the N-linked auxiliary functional group; andStep C: isolating the oligopeptide product having an amide bond that bears the N-linked auxiliary functional group.