Dynamic Conformational States Dictate Selectivity toward the Native Substrate in a Substrate-Permissive Acyltransferase

Biochemistry. 2016 Nov 15;55(45):6314-6326. doi: 10.1021/acs.biochem.6b00887. Epub 2016 Nov 2.

Abstract

Hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyltransferase (HCT) is an essential acyltransferase that mediates flux through plant phenylpropanoid metabolism by catalyzing a reaction between p-coumaroyl-CoA and shikimate, yet it also exhibits broad substrate permissiveness in vitro. How do enzymes like HCT avoid functional derailment by cellular metabolites that qualify as non-native substrates? Here, we combine X-ray crystallography and molecular dynamics to reveal distinct dynamic modes of HCT under native and non-native catalysis. We find that essential electrostatic and hydrogen-bonding interactions between the ligand and active site residues, permitted by active site plasticity, are elicited more effectively by shikimate than by other non-native substrates. This work provides a structural basis for how dynamic conformational states of HCT favor native over non-native catalysis by reducing the number of futile encounters between the enzyme and shikimate.

MeSH terms

  • Acyltransferases / chemistry*
  • Acyltransferases / genetics
  • Acyltransferases / metabolism
  • Amino Acid Sequence
  • Arabidopsis Proteins / chemistry*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Binding Sites / genetics
  • Biocatalysis
  • Catalytic Domain*
  • Crystallography, X-Ray
  • Hydrogen Bonding
  • Kinetics
  • Molecular Dynamics Simulation
  • Mutation
  • Protein Conformation*
  • Sequence Homology, Amino Acid
  • Shikimic Acid / chemistry
  • Shikimic Acid / metabolism
  • Static Electricity
  • Substrate Specificity

Substances

  • Arabidopsis Proteins
  • Shikimic Acid
  • Acyltransferases
  • HCT protein, Arabidopsis