Chemical shift mapping of shikimate‐3‐phosphate binding to the isolated N‐terminal domain of 5‐enolpyruvylshikimate‐3‐phosphate synthase

FEBS Letters

Young, John K.¹  Helms, Gregory L.²  Stauffer, Melissa E.¹  Evans, Jeremy N.S.¹ 

[1] School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USA;Center for NMR Spectroscopy, Washington State University, Pullman, WA 99164-4660, USA

关键词: Chemical shift mapping;    EPSP synthase;    NMR;    Secondary structure;    CSI;    chemical shift index;    EPSP;    5-enolpyruvylshikimate-3-phosphate;    GLP;    glyphosate;    NOE;    nuclear Overhauser effect;    S3P;    shikimate-3-phosphate;   

DOI  :  10.1016/S0014-5793(01)02555-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To facilitate evaluation of enzyme–ligand complexes in solution, we have isolated the 26-kDa N-terminal domain of 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase for analysis by NMR spectroscopy. The isolated domain is capable of binding the substrate shikimate-3-phosphate (S3P), and this letter reports the localization of the S3P binding site using chemical shift mapping. Based on the NMR data, we propose that Ser23, Arg27, Ser197, and Tyr200 are directly involved in S3P binding. We also describe changes in the observed nuclear Overhauser effects (NOEs) that are consistent with a partial conformational change in the N-terminal domain upon S3P binding.

【 授权许可】

Unknown