title: The site-specific incorporation of p-iodo-L-phenylalanine into proteins for structure determination.
abstract: A recently developed method makes it possible to genetically encode unnatural amino acids with diverse physical, chemical or biological properties in Escherichia coli and yeast. We now show that this technology can be used to efficiently and site-specifically incorporate p-iodo-L-phenylalanine (iodoPhe) into proteins in response to an amber TAG codon. The selective introduction of the anomalously scattering iodine atom into proteins should facilitate single-wavelength anomalous dispersion experiments on in-house X-ray sources. To illustrate this, we generated a Phe153 --> iodoPhe mutant of bacteriophage T4 lysozyme and determined its crystal structure using considerably less data than are needed for the equivalent experiment with cysteine and methionine. The iodoPhe residue, although present in the hydrophobic core of the protein, did not perturb the protein structure in any meaningful way. The ability to selectively introduce this and other heavy atom-containing amino acids into proteins should facilitate the structural study of proteins.
authors.forename: Jianming
authors.initials: J
authors.lastname: Xie
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Ning
authors.initials: N
authors.lastname: Wu
authors.forename: Ansgar
authors.initials: A
authors.lastname: Brock
authors.forename: Glen
authors.initials: G
authors.lastname: Spraggon
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
mesh_terms: D019943:Amino Acid Substitution; D017122:Bacteriophage T4 / Q000201:enzymology; D018360:Crystallography, X-Ray / Q000379:methods*; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000737:chemistry* / Q000235:genetics; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*
chemical_list: D015335:Molecular Probes; D011506:Proteins; D010649:Phenylalanine; D009113:Muramidase; C049181:4-iodophenylalanine