title: Multiply labeling proteins for studies of folding and stability.
abstract: Fluorescence spectroscopy is a powerful method for monitoring protein folding in real-time with high resolution and sensitivity, but requires the site-specific introduction of labels into the protein. The ability to genetically incorporate unnatural amino acids (Uaas) allows for the efficient synthesis of fluorescently labeled proteins with minimally perturbing fluorophores. Here, we describe recent uses of labeled proteins in dynamic structure determination experiments and advances in unnatural amino acid incorporation for dual site-specific fluorescent labeling. The advent of increasingly sophisticated bioorthogonal chemistry reactions and the diversity of Uaas available for incorporation will greatly enable protein folding and stability studies.
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104, United States.
authors.forename: Conor M
authors.initials: CM
authors.lastname: Haney
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104, United States.
authors.forename: Rebecca F
authors.initials: RF
authors.lastname: Wissner
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104, United States. Electronic address: ejpetersson@sas.upenn.edu.
authors.forename: E James
authors.initials: EJ
authors.lastname: Petersson
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics; D000818:Animals; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D017510:Protein Folding; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D000596:Amino Acids; D005456:Fluorescent Dyes; D011506:Proteins