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Study of the Phosphoryl-Transfer Mechanism of Shikimate Kinase by NMR Spectroscopy

Lookup NU author(s): Paul Thompson, Professor Alastair Hawkins

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Abstract

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 H-1 and (PNMR)-P-31 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 -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.

Publication metadata

Author(s): Prado V, Lence E, Vallejo JA, Beceiro A, Thompson P, Hawkins AR, Gonzalez-Bello C

Publication type: Article

Publication status: Published

Journal: Chemistry: A European Journal

Year: 2016

Volume: 22

Issue: 8

Pages: 2758-2768

Print publication date: 01/02/2016

Online publication date: 21/01/2016

Acceptance date: 01/01/1900

ISSN (print): 0947-6539

ISSN (electronic): 1521-3765

Publisher: Wiley - V C H Verlag GmbH & Co. KGaA

DOI: 10.1002/chem.201504438

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