title: Design of protein congeners containing β-cyclopropylalanine.
abstract: The non-canonical amino acid (ncAA) analogue of methionine (Met), β-cyclopropylalanine (Cpa), was successfully incorporated into recombinant proteins expressed in Escherichia coli in a residue-specific manner. Proteins substituted in this way are congeners because they derive from the same gene sequence as the parent protein but contain a fraction of ncAAs. We have expressed congeners using parent and mutant gene sequences of various proteins (lipase, annexin A5, enhanced green fluorescent protein, and barstar) and found that Cpa incorporation is highly dependent on the protein sequence composition. These results indicate that the global amino acid composition of proteins might be a crucial parameter that influences the outcome of unnatural translation. In addition, we could also demonstrate that the chemical nature of the second residue could be essential for successful ncAA incorporation.
authors: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim, Germany. acevedor@kofo.mpg.de
authors: Carlos G
authors: CG
authors: Acevedo-Rocha
authors: Anna-Skrollan
authors: AS
authors: Geiermann
authors: Nediljko
authors: N
authors: Budisa
authors: Lars
authors: L
authors: Merkel
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D017304:Annexin A5 / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008049:Lipase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013058:Mass Spectrometry; D008715:Methionine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: D017304:Annexin A5; D001426:Bacterial Proteins; D004988:Ethers, Cyclic; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C043657:cyclopropanol; C057438:barstar protein, Bacillus amyloliquefaciens; D008715:Methionine; D008049:Lipase; D000409:Alanine