title: Expressed protein ligation for metalloprotein design and engineering.
abstract: Metalloproteins contain highly specialized metal-binding sites that are designed to accept specific metal ions to maintain correct function. Although many of the sites have been modified with success, the relative paucity of functional group availability within proteinogenic amino acids can sometimes leave open questions about specific functions of the metal binding ligands. Attaining a more thorough analysis of individual amino acid function within metalloproteins has been realized using expressed protein ligation (EPL). Here we describe our recent efforts using EPL to incorporate nonproteinogenic cysteine and methionine analogues into the type 1 copper site found in Pseudomonas aeruginosa azurin.
authors.affiliation: Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Clark
authors.forename: Wilfred A
authors.initials: WA
authors.lastname: van der Donk
authors.forename: Yi
authors.initials: Y
authors.lastname: Lu
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000502:physiology; D001400:Azurin / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D003300:Copper / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D047668:Inteins; D008024:Ligands; D008667:Metalloproteins / Q000737:chemistry*; D008715:Methionine / Q000031:analogs & derivatives / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis*; D015202:Protein Engineering / Q000379:methods*; D019154:Protein Splicing; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017279:Selenocysteine / Q000031:analogs & derivatives / Q000737:chemistry; D013053:Spectrophotometry
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D008024:Ligands; D008667:Metalloproteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D017279:Selenocysteine; D001400:Azurin; D003300:Copper; D008715:Methionine; D003545:Cysteine