RT Journal Article
T1 Study of the Phosphoryl‐Transfer Mechanism of Shikimate Kinase by NMR Spectroscopy
A1 Prado, Verónica
A1 Lence Quintana, Emilio José
A1 Vallejo Vidal, Juan Andrés
A1 Beceiro Casas, Alejandro
A1 Thompson, Paul
A1 Hawkins, Alastair R.
A1 González Bello, Concepción
K1 Drug discovery
K1 Enzyme catalysis
K1 Molecular dynamics
K1 Molecular modelling
K1 Reaction mechanism
AB The 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 enzymes
PB Wiley
YR 2016
FD 2016-01-21
LK http://hdl.handle.net/10347/16937
UL http://hdl.handle.net/10347/16937
LA eng
NO 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. doi: 10.1002/chem.201504438
NO This 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-Archiving
NO Spanish Ministry of Economy and Competiveness. Grant Number: SAF2013-42899-RXunta de Galicia. Grant Number: GRC2013-041European Regional Development FundSara Borrell Programme. Grant Number: CD13/00373ISCIII General Subdirection of Assesment and Promotion of the Research. Grant Number: PI14/00059
DS Minerva
RD 1 may 2026