PDBsum entry: 2qfq

Transferase

PDB id: 2qfq

Contents

Protein chain

427 a.a. *

Ligands

S3P

FMT ×10

Waters ×561

* Residue conservation analysis

PDB id:

2qfq

Name:

Transferase

Title:

E. Coli epsp synthase pro101leu liganded with s3p

Structure:

3-phosphoshikimate 1-carboxyvinyltransferase. Chain: a. Synonym: 5-enolpyruvylshikimate-3-phosphate synthase, epsp epsps. Engineered: yes. Mutation: yes

Source:

Escherichia coli. Organism_taxid: 83333. Strain: k12. Gene: aroa. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

1.50Å R-factor: 0.164 R-free: 0.182

Authors:

E.Schonbrunn,M.L.Healy-Fried

Key ref:

M.L.Healy-Fried et al. (2007). Structural basis of glyphosate tolerance resulting from mutations of Pro101 in Escherichia coli 5-enolpyruvylshikimate-3-phosphate synthase. J Biol Chem, 282, 32949-32955. PubMed id: 17855366 DOI: 10.1074/jbc.M705624200

Date:

27-Jun-07 Release date: 02-Oct-07

Protein chain

P0A6D3  (AROA_ECOLI) -  3-phosphoshikimate 1-carboxyvinyltransferase

Seq: 427 a.a.

Struc: 427 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.5.1.19 - 3-phosphoshikimate 1-carboxyvinyltransferase.
• Pathway: Shikimate and Chorismate Biosynthesis
• Reaction: Phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O- (1-carboxyvinyl)-3-phosphoshikimate

Phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O- (1-carboxyvinyl)-3-phosphoshikimate (Bound ligand (Het Group name = S3P) matches with 76.19% similarity)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

 Gene Ontology (GO) functional annotation 

• Cellular component: cytoplasm (1 term)
• Biological process: cellular amino acid biosynthetic process (2 terms)
• Biochemical function: catalytic activity (4 terms)

reference

DOI no: 10.1074/jbc.M705624200

J Biol Chem 282:32949-32955 (2007)

PubMed id: 17855366

Structural basis of glyphosate tolerance resulting from mutations of Pro101 in Escherichia coli 5-enolpyruvylshikimate-3-phosphate synthase.

M.L.Healy-Fried, T.Funke, M.A.Priestman, H.Han, E.Schönbrunn.

ABSTRACT

Glyphosate, the world's most used herbicide, is a massive success because it enables efficient weed control with minimal animal and environmental toxicity. The molecular target of glyphosate is 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), which catalyzes the sixth step of the shikimate pathway in plants and microorganisms. Glyphosate-tolerant variants of EPSPS constitute the basis of genetically engineered herbicide-tolerant crops. A single-site mutation of Pro(101) in EPSPS (numbering according to the enzyme from Escherichia coli) has been implicated in glyphosate-resistant weeds, but this residue is not directly involved in glyphosate binding, and the basis for this phenomenon has remained unclear in the absence of further kinetic and structural characterization. To probe the effects of mutations at this site, E. coli EPSPS enzymes were produced with glycine, alanine, serine, or leucine substituted for Pro(101). These mutant enzymes were analyzed by steady-state kinetics, and the crystal structures of the substrate binary and substrate.glyphosate ternary complexes of P101S and P101L EPSPS were determined to between 1.5- and 1.6-A resolution. It appears that residues smaller than leucine may be substituted for Pro(101) without decreasing catalytic efficiency. Any mutation at this site results in a structural change in the glyphosate-binding site, shifting Thr(97) and Gly(96) toward the inhibitor molecule. We conclude that the decreased inhibitory potency observed for glyphosate is a result of these mutation-induced long-range structural changes. The implications of our findings concerning the development and spread of glyphosate-resistant weeds are discussed.

Selected figure(s)

Figure 1.
FIGURE 1. Reaction catalyzed by EPSPS. PEP, phosphoenolpyruvate.

Figure 2.
FIGURE 2. Location of Pro^101 in the structure of WT EPSPS from E. coli (stereo view). EPSPS is composed of two globular domains that close upon binding of S3P and glyphosate (shown in yellow and green, respectively); the two ligands are located in the interdomain cleft of the closed enzyme state (Protein Data Bank code 1g6s) (6). Displayed in maroon is the helix in the upper (N-terminal) domain containing Pro^101. Glyphosate binds adjacent to S3P, its phosphonate moiety pointing toward the N-terminal end of the helix.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 32949-32955) copyright 2007.

Figures were selected by the author.