title: Intracellular quantitative detection of human thymidylate synthase engagement with an unconventional inhibitor using tetracysteine-diarsenical-probe technology.
abstract: Demonstrating a candidate drug's interaction with its target protein in live cells is of pivotal relevance to the successful outcome of the drug discovery process. Although thymidylate synthase (hTS) is an important anticancer target protein, the efficacy of the few anti-hTS drugs currently used in clinical practice is limited by the development of resistance. Hence, there is an intense search for new, unconventional anti-hTS drugs; there are approximately 1600 ongoing clinical trials involving hTS-targeting drugs, both alone and in combination protocols. We recently discovered new, unconventional peptidic inhibitors of hTS that are active against cancer cells and do not result in the overexpression of hTS, which is a known molecular source of resistance. Here, we propose an adaptation of the recently proposed tetracysteine-arsenic-binding-motif technology to detect and quantitatively characterize the engagement of hTS with one such peptidic inhibitor in cell lysates. This new model can be developed into a test for high-throughput screening studies of intracellular target-protein/small-molecule binding.
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.
authors.forename: Glauco
authors.initials: G
authors.lastname: Ponterini
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.|University of Edinburgh, University/British Heart Foundation Centre for Cardiovascular Science, The Queen's Medical Research Institute, Edinburgh, EH16 4TJ, UK.
authors.forename: Andrea
authors.initials: A
authors.lastname: Martello
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Pavesi
authors.affiliation: University of Modena and Reggio Emilia, Department of Biomedical, Metabolic and Neural Sciences, Via Giuseppe Campi 287, Modena, 41125, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Lauriola
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Luciani
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Santucci
authors.affiliation: University of Ferrara, Department of Chemical and Pharmaceutical Sciences, Via Fossato di Mortara 17-19, Ferrara, 44100, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Pelà
authors.affiliation: University of Modena and Reggio Emilia, Department of Biomedical, Metabolic and Neural Sciences, Via Giuseppe Campi 287, Modena, 41125, Italy.
authors.forename: Gaia
authors.initials: G
authors.lastname: Gozzi
authors.affiliation: University of Ferrara, Department of Chemical and Pharmaceutical Sciences, Via Fossato di Mortara 17-19, Ferrara, 44100, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Pacifico
authors.affiliation: University of Ferrara, Department of Chemical and Pharmaceutical Sciences, Via Fossato di Mortara 17-19, Ferrara, 44100, Italy.
authors.forename: Remo
authors.initials: R
authors.lastname: Guerrini
authors.affiliation: University of Modena and Reggio Emilia, Department of Biomedical, Metabolic and Neural Sciences, Via Giuseppe Campi 287, Modena, 41125, Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Marverti
authors.affiliation: University of Modena and Reggio Emilia, Department of Life Sciences, Via Giuseppe Campi 183, Modena, 41125, Italy.
authors.forename: Maria Paola
authors.initials: MP
authors.lastname: Costi
authors.affiliation: University of Modena and Reggio Emilia, Department of Biomedical, Metabolic and Neural Sciences, Via Giuseppe Campi 287, Modena, 41125, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: D'Arca
mesh_terms: D020816:Amino Acid Motifs; D001151:Arsenic / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain; D003545:Cysteine / Q000737:chemistry* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D013940:Thymidylate Synthase; D003545:Cysteine; D001151:Arsenic