Prado, VerónicaLence Quintana, Emilio JoséVallejo Vidal, Juan AndrésBeceiro Casas, AlejandroThompson, PaulHawkins, Alastair R.González Bello, Concepción2018-07-032018-07-032016-01-21Prado, V., Lence, E., Vallejo, J., Beceiro, A., Thompson, P., Hawkins, A., & González-Bello, C. (2016). Study of the Phosphoryl-Transfer Mechanism of Shikimate Kinase by NMR Spectroscopy. Chemistry - A European Journal, 22(8), 2758-2768. doi: 10.1002/chem.201504438http://hdl.handle.net/10347/16937This is the peer-reviewed version of the following article: Prado, V., Lence, E., Vallejo, J., Beceiro, A., Thompson, P., Hawkins, A., & González-Bello, C. (2016). Study of the Phosphoryl-Transfer Mechanism of Shikimate Kinase by NMR Spectroscopy. Chemistry - A European Journal, 22(8), 2758-2768, which has been published in final form at https://doi.org/10.1002/chem.201504438. This article may be used for non-commercial purposes in accordance with Wiley-VCH Terms and Conditions for Self-ArchivingThe phosphoryl‐transfer mechanism of shikimate kinase from Mycobacterium tuberculosis and Helicobacter pylori, which is an attractive target for antibiotic drug discovery, has been studied by 1D 1H and 31P NMR spectroscopy. Metaphosphoric acid proved to be a good mimetic of the metaphosphate intermediate and facilitated the ready and rapid evaluation by NMR spectroscopic analysis of a dissociative mechanism. The required closed form of the active site for catalysis was achieved by the use of ADP (product) or two synthetic ADP analogues (AMPNP, AMPCP). Molecular dynamics simulation studies reported here also revealed that the essential arginine (Arg116/Arg117 in H. pylori and M. tuberculosis, respectively), which activates the γ‐phosphate group of ATP for catalysis and triggers the release of the product for turnover, would also be involved in the stabilisation of the metaphosphate intermediate during catalysis. We believe that the studies reported here will be helpful for future structure‐based design of inhibitors of this attractive target. The approach is also expected be useful for studies on the possible dissociative mechanism of other kinase enzymeseng© 2016 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. This article may be used for non-commercial purposes in accordance with Wiley-VCH Terms and Conditions for Self-ArchivingDrug discoveryEnzyme catalysisMolecular dynamicsMolecular modellingReaction mechanismjournal article10.1002/chem.2015044381521-3765open access