Nature Biotechnology22, 1297 - 1301 (2004)
Published online: 19 September 2004; | doi:10.1038/nbt1013
The site-specific incorporation of p-iodo-L-phenylalanine into proteins for structure determination
Jianming Xie1, Lei Wang1, Ning Wu1, Ansgar Brock2, Glen Spraggon2
& Peter G Schultz1, 2
1
Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
2
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, USA.
A recently developed method makes it possible to genetically encode unnatural amino acids with diverse physical, chemical or biological properties in Escherichia coli1 and yeast2. 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 dispersion3,
4 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.
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
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.
