tailieunhanh - Báo cáo khoa học: Nucleotide binding to human UMP-CMP kinase using fluorescent derivatives ) a screening based on affinity for the UMP-CMP binding site

Methylanthraniloyl derivatives of ATP and CDP were usedin 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. | ễFEBS Journal Nucleotide binding to human UMP-CMP kinase using fluorescent derivatives - a screening based on affinity for the UMP-CMP binding site Dimitri Topalis1 Hiroki Kumamoto2 Maria-Fernanda Amaya Velasco3 Laurence Dugue4 Ahmed Haouz5 Julie Anne C. Alexandre1 Sarah Gallois-Montbrun6 Pedro Maria Alzari3 Sylvie Pochet4 Luigi Andre Agrofoglio2 and Dominique Deville-Bonne1 1 Laboratoire d Enzymologie Moleculaire et Fonctionnelle FRE 2852 CNRS-Paris 6 Institut Jacques Monod Paris France 2 Institut de Chimie Organique et Analytique UMR CNRS 6005 FR 2708 Universite d Orleans UFR Sciences Orleans France 3 Unite de Biochimie Structurale URA CNRS 2185 Institut Pasteur Paris France 4 Unite de Chimie Organique URA CNRS 2128 Institut Pasteur Paris France 5 Plate-Forme 6- Cristallogenese et Diffraction des Rayons X Institut Pasteur Paris France 6 Unite de Regulation Enzymatique des Activites Cellulaires CNRS URA 2185 Institut Pasteur Paris France Keywords cidofovir human UMP-CMP kinase MABA-CDP Mant-ATP phosphonates Correspondence D. Deville-Bonne Laboratoire d Enzymologie Moleculaire et Fonctionnelle FRE 2852 CNRS-Paris 6 Institut Jacques Monod 4 place Jussieu 75251 Paris Cedex 05 France Fax 33 1 44 27 59 94 Tel 33 1 44 27 59 93 E-mail ddeville@ These authors contributed equally to this work Present address Department of Infectious Diseases Guy s King s and St Thomas Medical School King s College London GKT Guy s Hospital London Received 12 February 2007 revised 25 May 2007 accepted 29 May 2007 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 .