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Nitric Oxide Sensing by a Blue Fluorescent Protein.
Montali, Chiara; Abbruzzetti, Stefania; Franzen, Arne; Casini, Giorgia; Bruno, Stefano; Delcanale, Pietro; Burgstaller, Sandra; Ramadani-Muja, Jeta; Malli, Roland; Gensch, Thomas; Viappiani, Cristiano.
Afiliação
  • Montali C; Dipartimento di Scienze Matematiche, Fisiche e Informatiche, Università di Parma, Parco Area delle Scienze 7A, 43124 Parma, Italy.
  • Abbruzzetti S; Dipartimento di Scienze Matematiche, Fisiche e Informatiche, Università di Parma, Parco Area delle Scienze 7A, 43124 Parma, Italy.
  • Franzen A; Institute of Biological Information Processing (IBI-1: Molecular and Cellular Physiology), Forschungszentrum Jülich, Leo-Brandt-Straße, D-52428 Jülich, Germany.
  • Casini G; Institute of Biological Information Processing (IBI-1: Molecular and Cellular Physiology), Forschungszentrum Jülich, Leo-Brandt-Straße, D-52428 Jülich, Germany.
  • Bruno S; Dipartimento di Scienze degli Alimenti e del Farmaco, Università di Parma, Parco Area delle Scienze 23/A, 43124 Parma, Italy.
  • Delcanale P; Dipartimento di Scienze Matematiche, Fisiche e Informatiche, Università di Parma, Parco Area delle Scienze 7A, 43124 Parma, Italy.
  • Burgstaller S; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Neue Stiftingtalstraße 6/6, 8010 Graz, Austria.
  • Ramadani-Muja J; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Neue Stiftingtalstraße 6/6, 8010 Graz, Austria.
  • Malli R; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Neue Stiftingtalstraße 6/6, 8010 Graz, Austria.
  • Gensch T; Institute of Biological Information Processing (IBI-1: Molecular and Cellular Physiology), Forschungszentrum Jülich, Leo-Brandt-Straße, D-52428 Jülich, Germany.
  • Viappiani C; Dipartimento di Scienze Matematiche, Fisiche e Informatiche, Università di Parma, Parco Area delle Scienze 7A, 43124 Parma, Italy.
Antioxidants (Basel) ; 11(11)2022 Nov 11.
Article em En | MEDLINE | ID: mdl-36421416
S-Nitrosylation of cysteine residues is an important molecular mechanism for dynamic, post-translational regulation of several proteins, providing a ubiquitous redox regulation. Cys residues are present in several fluorescent proteins (FP), including members of the family of Aequorea victoria Green Fluorescent Protein (GFP)-derived FPs, where two highly conserved cysteine residues contribute to a favorable environment for the autocatalytic chromophore formation reaction. The effect of nitric oxide on the fluorescence properties of FPs has not been investigated thus far, despite the tremendous role FPs have played for 25 years as tools in cell biology. We have examined the response to nitric oxide of fluorescence emission by the blue-emitting fluorescent protein mTagBFP2. To our surprise, upon exposure to micromolar concentrations of nitric oxide, we observed a roughly 30% reduction in fluorescence quantum yield and lifetime. Recovery of fluorescence emission is observed after treatment with Na-dithionite. Experiments on related fluorescent proteins from different families show similar nitric oxide sensitivity of their fluorescence. We correlate the effect with S-nitrosylation of Cys residues. Mutation of Cys residues in mTagBFP2 removes its nitric oxide sensitivity. Similarly, fluorescent proteins devoid of Cys residues are insensitive to nitric oxide. We finally show that mTagBFP2 can sense exogenously generated nitric oxide when expressed in a living mammalian cell. We propose mTagBFP2 as the starting point for a new class of genetically encoded nitric oxide sensors based on fluorescence lifetime imaging.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Antioxidants (Basel) Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Itália País de publicação: Suíça

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Antioxidants (Basel) Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Itália País de publicação: Suíça