| PMID : |
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 (17608725) |
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| titre : |
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Nucleotide binding to human UMP-CMP kinase using fluorescent derivatives - a screening based on affinity for the UMP-CMP binding site. |
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| auteur(s) : |
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Dimitri Topalis1, Hiroki Kumamoto2, Maria-Fernanda Amaya Velasco3, Laurence Dugué4, Ahmed Haouz5, Julie Anne C Alexandre1, Sarah Gallois-Montbrun6, Pedro Maria Alzari3, Sylvie Pochet4, Luigi André Agrofoglio2, Dominique Deville-Bonne1 |
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| laboratoire : |
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| résumé : |
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Methylanthraniloyl derivatives of ATP and CDP were used in vitro as fluorescent probes for the donor-binding and acceptor-binding sites of human UMP-CMP kinase, a nucleoside salvage pathway kinase. Like all NMP kinases, UMP-CMP kinase binds the phosphodonor, usually ATP, and the NMP at different binding sites. The reaction results from an in-line phosphotransfer from the donor to the acceptor. The probe for the donor site was displaced by the bisubstrate analogs of the Ap5X series (where X = U, dT, A, G), indicating the broad specificity of the acceptor site. Both CMP and dCMP were competitors for the acceptor site probe. To find antimetabolites for antivirus and anticancer therapies, we have developed a method of screening acyclic phosphonate analogs that is based on the affinity of the acceptor-binding site of the human UMP-CMP kinase. Several uracil vinylphosphonate derivatives had affinities for human UMP-CMP kinase similar to those of dUMP and dCMP and better than that of cidofovir, an acyclic nucleoside phosphonate with a broad spectrum of antiviral activities. The uracil derivatives were inhibitors rather than substrates of human UMP-CMP kinase. Also, the 5-halogen-substituted analogs inhibited the human TMP kinase less efficiently. The broad specificity of the enzyme acceptor-binding site is in agreement with a large substrate-binding pocket, as shown by the 2.1 A crystal structure. |
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| domaine : |
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Sciences du Vivant/Autre
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langue du texte
intégral : |
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Anglais |
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| ISSN : |
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1742-464X |
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| type de publication : |
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Articles dans des revues avec comité de lecture |
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| DOI : |
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10.1111/j.1742-4658.2007.05902.x |
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| journal : |
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The FEBS Journal |
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| Audience : |
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non spécifiée |
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| date de publication : |
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07/2007 |
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| volume : |
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274 |
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| numéro : |
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14 |
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| page, identifiant, ... : |
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3704-14 |
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