title: Physical and structural properties of [Cu(BOT1)Cl]Cl, a fluorescent imaging probe for HNO.
abstract: Nitroxyl, or HNO, is involved in a number of important physiological processes, such as vascular relaxation and neuroregulation. Effective imaging tools are required in order to gain a deeper understanding of the in vivo mechanisms of these processes and to identify the endogenous sources of HNO. Here, we further investigate the physical properties of our previously reported fluorescent nitroxyl sensor, [Cu(BOT1)Cl]Cl (J. Am. Chem. Soc.2010, 132, 5536; BOT1=BODIPY·triazole, a tetradentate ligand). A new high-yielding synthetic procedure for BOT1 is reported. The X-ray crystal structures of two Cu(II) complexes of BOT1 are described. These structural studies show that the BOT1 ligand can form Cu(II) coordination complexes of both square-pyramidal and trigonal-bipyramidal geometries. Cyclic voltammograms of [Cu(BOT1)Cl]Cl were acquired, revealing the presence of a quasi-reversible feature at 130 mV (vs the ferrocene/ferrocenium couple) in MeCN and at -40 mV (vs Ag/AgCl) in aqueous buffer, which is assigned to the Cu(II)/Cu(I) couple. The reactivity of [Cu(BOT1)Cl]Cl with Angeli's salt, a stable source of HNO, was further investigated. A 1000-fold excess of Angeli's salt elicits an immediate 10-fold emission turn-on response of the sensor, consistent with our previous report. A new observation, reported here, is that the intensity of this turn-on emission diminishes at longer incubation times. Fluorescent imaging of nitroxyl by [Cu(BOT1)Cl]Cl in HeLa cells was carried out. Upon treatment of the cells with Angeli's salt, there was a modest 2-fold intracellular turn-on in emission intensity.
authors.affiliation: Massachusetts Institute of Technology, Department of Chemistry, Cambridge, MA 02139, USA.
authors.forename: Maksim
authors.initials: M
authors.lastname: Royzen
authors.forename: Justin J
authors.initials: JJ
authors.lastname: Wilson
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Lippard
mesh_terms: D003300:Copper / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004563:Electrochemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D008958:Models, Molecular; D008968:Molecular Conformation; D009589:Nitrogen Oxides / Q000737:chemistry* / Q000378:metabolism; D009942:Organometallic Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction
chemical_list: D005456:Fluorescent Dyes; D009589:Nitrogen Oxides; D009942:Organometallic Compounds; D003300:Copper; C039900:nitroxyl