title: Semi-synthesis of labeled proteins for spectroscopic applications.
abstract: Since the introduction of SPPS by Merrifield in the 60s, peptide chemists have considered the possibility of preparing large proteins. The introduction of native chemical ligation in the 90s and then of expressed protein ligation have opened the way to the preparation of synthetic proteins without size limitations. This review focuses on semi-synthetic strategies useful to prepare proteins decorated with spectroscopic probes, like fluorescent labels and stable isotopes, and their biophysical applications. We show that expressed protein ligation, combining the advantages of organic chemistry with the easy and size limitless recombinant protein expression, is an excellent strategy for the chemical synthesis of labeled proteins, enabling a single protein to be functionalized at one or even more distinct positions with different probes.
authors.affiliation: Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, Napoli 80134, Italy. ldandrea@unina.it
authors.forename: Lucia
authors.initials: L
authors.lastname: De Rosa
authors.forename: Anna
authors.initials: A
authors.lastname: Russomanno
authors.forename: Alessandra
authors.initials: A
authors.lastname: Romanelli
authors.forename: Luca Domenico
authors.initials: LD
authors.lastname: D'Andrea
mesh_terms: D020816:Amino Acid Motifs; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D047668:Inteins; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering; D011506:Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D010446:Peptide Fragments; D011506:Proteins; D011994:Recombinant Proteins