title: Chemical synthesis of cell-permeable apoptotic peptides from in vivo produced proteins.
abstract: In vivo synthesis of peptides by bacterial expression has developed into a reliable alternative to solid-phase peptide synthesis. A significant drawback of in vivo methods is the difficulty with which gene products can be modified post-translationally. Here, we present a method for the facile modification of peptides generated in bacterial hosts after cyanogen bromide cleavage at C-terminal methionines. Reaction of the resulting homoserine lactones with propargylamine allows efficient and selective modification with a wide variety of chemicals such as fluorescent dyes, biotin derivatives, polyprenyls, lipids, polysaccharides, or peptides. Attachment of the cell penetrating peptide octa-arginine (R(8)) to peptides derived from the proapoptotic tumor suppressor Bak BH3 led to efficient cellular uptake and subsequent cytochrome c release from mitochondria, culminating in induction of apoptosis similar to that observed with peptides linked to R(8) via the peptide backbone. These results highlight the significant potential for use of such tools in live cells.
authors.affiliation: School of Chemistry, Cardiff University, Park Place, Cardiff, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Fricke
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Mart
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Watkins
authors.forename: Marie
authors.initials: M
authors.lastname: Wiltshire
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Errington
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Smith
authors.forename: Arwyn T
authors.initials: AT
authors.lastname: Jones
authors.forename: Rudolf K
authors.initials: RK
authors.lastname: Allemann
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000737:chemistry; D000595:Amino Acid Sequence; D017209:Apoptosis; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D003488:Cyanogen Bromide / Q000737:chemistry; D045304:Cytochromes c / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis* / Q000493:pharmacokinetics*; D015202:Protein Engineering / Q000379:methods*; D011518:Proto-Oncogene Proteins / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000737:chemistry
chemical_list: C474007:Bax protein (53-86); D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D050998:bcl-2 Homologous Antagonist-Killer Protein; C500307:octa-arginine peptide; C088386:homoserine lactone; D045304:Cytochromes c; D015107:4-Butyrolactone; D003488:Cyanogen Bromide