title: Designed to dissolve: suppression of colloidal aggregation of Cu(I)-selective fluorescent probes in aqueous buffer and in-gel detection of a metallochaperone.
abstract: Due to the lipophilicity of the metal-ion receptor, previously reported Cu(I)-selective fluorescent probes form colloidal aggregates, as revealed by dynamic light scattering. To address this problem, we have developed a hydrophilic triarylpyrazoline-based fluorescent probe, CTAP-2, that dissolves directly in water and shows a rapid, reversible, and highly selective 65-fold fluorescence turn-on response to Cu(I) in aqueous solution. CTAP-2 proved to be sufficiently sensitive for direct in-gel detection of Cu(I) bound to the metallochaperone Atox1, demonstrating the potential for cation-selective fluorescent probes to serve as tools in metalloproteomics for identifying proteins with readily accessible metal-binding sites.
authors.affiliation: School of Chemistry and Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive, Atlanta, Georgia 30332, United States.
authors.forename: M Thomas
authors.initials: MT
authors.lastname: Morgan
authors.forename: Pritha
authors.initials: P
authors.lastname: Bagchi
authors.forename: Christoph J
authors.initials: CJ
authors.lastname: Fahrni
mesh_terms: D027682:Cation Transport Proteins / Q000032:analysis*; D003102:Colloids / Q000737:chemistry; D003300:Copper / Q000737:chemistry*; D000079922:Copper Transport Proteins; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D056511:Metallochaperones; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000032:analysis*; D011720:Pyrazoles / Q000737:chemistry; D012995:Solubility; D014867:Water / Q000737:chemistry
chemical_list: C474500:ATOX1 protein, human; D027682:Cation Transport Proteins; D003102:Colloids; D000079922:Copper Transport Proteins; D005456:Fluorescent Dyes; D056511:Metallochaperones; D018832:Molecular Chaperones; D011720:Pyrazoles; D014867:Water; D003300:Copper