Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3-Phosphoshikimate 1-Carboxyvinyltransferase
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Alkyl and Aryl Transferases / chemistry*
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Alkyl and Aryl Transferases / metabolism*
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Amino Acid Sequence
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Binding Sites
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Models, Molecular
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Molecular Sequence Data
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Nuclear Magnetic Resonance, Biomolecular
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Protein Binding
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Shikimic Acid / analogs & derivatives*
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Shikimic Acid / chemistry
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Shikimic Acid / metabolism*
Substances
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Shikimic Acid
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shikimic acid-3-phosphate
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Alkyl and Aryl Transferases
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3-Phosphoshikimate 1-Carboxyvinyltransferase