title: Native chemical ligation derived method for recombinant peptide/protein C-terminal amidation.
abstract: C-terminal amidation is often a requisite structural feature for peptide hormone bio-activity. We report a chemical amidation method that converts peptide/protein thioesters into their corresponding C-terminal amides. The peptide/protein thioester is treated with 1-(2,4-dimethoxyphenyl)-2-mercaptoethyl auxiliary (1b) in a native chemical ligation (NCL) reaction to form an intermediate, which upon removal of the auxiliary with TFA, yields the peptide/protein amide. We have demonstrated the general utility of the approach by successfully converting several synthetic peptide thioesters to peptide amides with high conversion rates. Preliminary results of converting a recombinant peptide thioester to its amide form are also reported.
authors.affiliation: Amylin Pharmaceuticals, LLC, San Diego, CA 92121, USA. chengzao.sun@amylin.com
authors.forename: Chengzao
authors.initials: C
authors.lastname: Sun
authors.forename: Gary
authors.initials: G
authors.lastname: Luo
authors.forename: Swetha
authors.initials: S
authors.lastname: Neravetla
authors.forename: Soumitra S
authors.initials: SS
authors.lastname: Ghosh
authors.forename: Bruce
authors.initials: B
authors.lastname: Forood
mesh_terms: D000577:Amides / Q000138:chemical synthesis* / Q000737:chemistry; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry*; D011486:Protein C / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D000577:Amides; D010455:Peptides; D011486:Protein C; D011994:Recombinant Proteins
keywords: 1-Phenyl-2-mercaptoethyl auxiliary; C-terminal amidation; Native chemical ligation; Peptide amide; Peptide thioester