title: Exploration of biarsenical chemistry--challenges in protein research.
abstract: The fluorescent modification of proteins (with genetically encoded low-molecular-mass fluorophores, affinity probes, or other chemically active species) is extraordinarily useful for monitoring and controlling protein functions in vitro, as well as in cell cultures and tissues. The large sizes of some fluorescent tags, such as fluorescent proteins, often perturb normal activity and localization of the protein of interest, as well as other effects. Of the many fluorescent-labeling strategies applied to in vitro and in vivo studies, one is very promising. This requires a very short (6- to 12-residue), appropriately spaced, tetracysteine sequence (-CCXXCC-); this is either placed at a protein terminus, within flexible loops, or incorporated into secondary structure elements. Proteins that contain the tetracysteine motif become highly fluorescent upon labeling with a nonluminescent biarsenical probe, and form very stable covalent complexes. We focus on the development, growth, and multiple applications of this protein research methodology, both in vitro and in vivo. Its application is not limited to intact-cell protein visualization; it has tremendous potential in other protein research disciplines, such as protein purification and activity control, electron microscopy imaging of cells or tissue, protein-protein interaction studies, protein stability, and aggregation studies.
authors.affiliation: Department of Protein Engineering, Faculty of Biotechnology, University of Wrocław, Wrocław, Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Pomorski
authors.forename: Artur
authors.initials: A
authors.lastname: Krężel
mesh_terms: D001151:Arsenic / Q000737:chemistry*; D002232:Carbocyanines / Q000737:chemistry; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D015394:Molecular Structure; D010078:Oxazines / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D010078:Oxazines; D011506:Proteins; C089187:cyanine dye 3; C014180:resorufin; D001151:Arsenic; D019793:Fluorescein