title: New Dual Fluorescent Probe for Simultaneous Biothiol and Phosphate Bioimaging.
abstract: The simultaneous detection of relevant metabolites in living organisms by using one molecule introduces an approach to understanding the relationships between these metabolites in healthy and deregulated cells. Fluorescent probes of low toxicity are remarkable tools for this type of analysis of biological systems in vivo. As a proof of concept, different naturally occurring compounds, such as biothiols and phosphate anions, were the focus for this work. The 2,4-dinitrobenzenesulfinate (DNBS) derivative of 9-[1-(4-tert-butyl-2-methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one (Granada Green; GG) were designed and synthesized. This new sulfinyl xanthene derivative can act as a dual sensor for the aforementioned analytes simultaneously. The mechanism of action of this derivative implies thiolysis of the sulfinyl group of the weakly fluorescent DNBS-GG by biological thiols at near-neutral pH values, thus releasing the fluorescent GG moiety, which simultaneously responds to phosphate anions through its fluorescence-decay time. The new dual probe was tested in solution by using steady-state and time-resolved fluorescence and intracellularly by using fluorescence-lifetime imaging microscopy (FLIM) in human epithelioid cervix carcinoma (HeLa) cells.
authors.affiliation: Department of Organic Chemistry, Faculty of Sciences, University of Granada, C. U. Fuentenueva s/n, 18071 Granada (Spain), Fax: (+34) 958248437.
authors.forename: Sandra
authors.initials: S
authors.lastname: Resa
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090.
authors.forename: Angel
authors.initials: A
authors.lastname: Orte
authors.affiliation: Department of Organic Chemistry, Faculty of Sciences, University of Granada, C. U. Fuentenueva s/n, 18071 Granada (Spain), Fax: (+34) 958248437.
authors.forename: Delia
authors.initials: D
authors.lastname: Miguel
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Paredes
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090.
authors.forename: Virginia
authors.initials: V
authors.lastname: Puente-Muñoz
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain).
authors.forename: Rafael
authors.initials: R
authors.lastname: Salto
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain).
authors.forename: Maria D
authors.initials: MD
authors.lastname: Giron
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Ruedas-Rama
authors.affiliation: Department of Organic Chemistry, Faculty of Sciences, University of Granada, C. U. Fuentenueva s/n, 18071 Granada (Spain), Fax: (+34) 958248437. jmcuerva@ugr.es.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Cuerva
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Alvarez-Pez
authors.affiliation: Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Cartuja Campus, 18071 Granada (Spain), Fax: (+34) 958244090. luiscrovetto@ugr.es.
authors.forename: Luis
authors.initials: L
authors.lastname: Crovetto
mesh_terms: D005260:Female; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009574:Nitro Compounds / Q000737:chemistry*; D010710:Phosphates / Q000032:analysis / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000737:chemistry*; D013452:Sulfonium Compounds / Q000737:chemistry*; D002583:Uterine Cervical Neoplasms / Q000737:chemistry*; D014966:Xanthenes / Q000737:chemistry*; D014970:Xanthines / Q000737:chemistry*
chemical_list: C000606953:2,4-dinitrobenzenesulfinate; C000606955:9-(1-(4-tert-butyl-2-methoxyphenyl))-6-hydroxy-3H-xanthen-3-one; D005456:Fluorescent Dyes; D009574:Nitro Compounds; D010710:Phosphates; D013438:Sulfhydryl Compounds; D013452:Sulfonium Compounds; D014966:Xanthenes; D014970:Xanthines
keywords: anions; fluorescence lifetime imaging microscopy; fluorescent probes; intracellular imaging agents; thiols