title: Designing novel spectral classes of proteins with a tryptophan-expanded genetic code.
abstract: Fluorescence methods are now well-established and powerful tools to study biological macromolecules. The canonical amino acid tryptophan (Trp), encoded by a single UGG triplet, is the main reporter of intrinsic fluorescence properties of most natural proteins and peptides and is thus an attractive target for tailoring their spectral properties. Recent advances in research have provided substantial evidence that the natural protein translational machinery can be genetically reprogrammed to introduce a large number of non-coded (i.e. noncanonical) Trp analogues and surrogates into various proteins. Especially attractive targets for such an engineering approach are fluorescent proteins in which the chromophore is formed post-translationally from an amino acid sequence, like the green fluorescent protein from Aequorea victoria. With the currently available translationally active fluoro-, hydroxy-, amino-, halogen-, and chalcogen-containing Trp analogues and surrogates, the traditional methods for protein engineering and design can be supplemented or even fully replaced by these novel approaches. Future research will provide a further increase in the number of Trp-like amino acids that are available for redesign (by engineering of the genetic code) of native Trp residues and enable novel strategies to generate proteins with tailored spectral properties.
authors.affiliation: Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, D-82152 Martinsried, Germany. budisa@biochem.mpg.de
authors.forename: Nediljko
authors.initials: N
authors.lastname: Budisa
authors.forename: Prajna Paramita
authors.initials: PP
authors.lastname: Pal
mesh_terms: D000818:Animals; D005815:Genetic Code; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics*; D014364:Tryptophan / Q000737:chemistry / Q000235:genetics*
chemical_list: D011506:Proteins; D014364:Tryptophan