title: Peptide chemical ligation inside living cells: in vivo generation of a circular protein domain.
abstract: Here we describe the first example of a peptide chemical ligation reaction performed inside a living cell. A cell-based native chemical ligation approach was developed and used to generate a circular version of the N-terminal Src homology 3 (SH3) domain from the murine c-Crk adapter protein inside Escherichia coli cells. The in vivo cyclization reaction was extremely efficient and the resulting circular protein domain was fully biologically active and able to adopt the native SH3 folded structure. This work represents an important step towards the in vivo generation of small backbone cyclic peptides for use in basic biological research.
authors.affiliation: Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, NY 10021, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Camarero
authors.forename: D
authors.initials: D
authors.lastname: Fushman
authors.forename: D
authors.initials: D
authors.lastname: Cowburn
authors.forename: T W
authors.initials: TW
authors.lastname: Muir
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003500:Cyclization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D063208:Methionyl Aminopeptidases; D051379:Mice; D010456:Peptides, Cyclic / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D018909:src Homology Domains / Q000235:genetics
chemical_list: D010456:Peptides, Cyclic; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases