Deciphering the structure of Arabidopsis thaliana 5-enol-pyruvyl-shikimate-3-phosphate synthase: An essential step toward the discovery of novel inhibitors to supersede glyphosate

Ruszkowski, Milosz; Forlani, Giuseppe

doi:10.1016/j.csbj.2022.03.020

Deciphering the structure of Arabidopsis thaliana 5-enol-pyruvyl-shikimate-3-phosphate synthase: An essential step toward the discovery of novel inhibitors to supersede glyphosate

Journal Article · Thu Mar 24 00:00:00 EDT 2022 · Computational and Structural Biotechnology Journal

DOI:https://doi.org/10.1016/j.csbj.2022.03.020· OSTI ID:1867474

Ruszkowski, Milosz ^[1]; Forlani, Giuseppe ^[2]

Polish Academy of Sciences (PAS), Poznan (Poland)
Univ. of Ferrara (Italy)

Glyphosate interferes with plant aromatic metabolism through the inhibition of 5-enol-pyruvyl-shikimate-3-phosphate (EPSP) synthase [EPSPS, EC 2.5.1.19]. For this reason, EPSPS has been extensively studied in a vast array of organisms. This notwithstanding, up to date, the crystal structure of the protein has been solved exclusively in a few prokaryotes, while that of the plant enzyme has been only deduced in silico by similarity. This study aimed at determining the structure of EPSPS from the plant model species Arabidopsis thaliana, which has been cloned, heterologously expressed and affinity-purified. The kinetic properties of the enzyme have been determined, as well as its susceptibility to the inhibition brought about by glyphosate. The crystal structure of the protein has been resolved at high resolution (1.4 Å), showing open conformation of the enzyme, which is the state ready for substrate/inhibitor binding. This provides a framework for the structure-based design of novel EPSPS inhibitors. Surface regions near the active-site cleft entrance or at the interdomain hinge appear promising for inhibitor selectivity, while bound chloride near the active site is a potential placeholder for anionic moieties of future herbicides.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Science Centre

Grant/Contract Number:: W-31109-ENG-38

OSTI ID:: 1867474

Journal Information:: Computational and Structural Biotechnology Journal, Journal Name: Computational and Structural Biotechnology Journal Vol. 20; ISSN 2001-0370

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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