Crystal structure of apo-glycolate oxidase

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T Sandalova1, Y Lindqvist

  • 1Department of Molecular Biology, Swedish University of Agricultural Sciences, Uppsala.

FEBS Letters
|August 2, 1993
PubMed

Insights

The crystal structure of apo-glycolate oxidase reveals significant flexibility and altered subunit conformations after cofactor removal. This apoform adopts a molten globule-like state, impacting protein structure and loop arrangements.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Enzymology

Background:

  • Glycolate oxidase is an alpha/beta-barrel enzyme crucial in metabolic pathways.
  • Cofactor binding, specifically flavin mononucleotide (FMN), is essential for enzyme activity.
  • Understanding apoenzyme structure provides insights into conformational changes upon cofactor interaction.

Purpose of the Study:

  • To determine the crystal structure of the apoform of glycolate oxidase.
  • To investigate the structural impact of flavin mononucleotide (FMN) removal on the enzyme.
  • To characterize the conformational changes associated with the loss of the tightly bound cofactor.

Main Methods:

  • X-ray crystallography was employed to resolve the protein structure.
  • The crystal structure was determined to a resolution of 2.6 Å.
  • Comparative analysis was performed between the apoenzyme and holoenzyme structures.

Main Results:

  • The apoform of glycolate oxidase exhibits a highly flexible, molten globule-like state.
  • The asymmetric unit revealed two subunits with distinct conformations compared to each other and the holoenzyme.
  • While secondary structures remained intact, their arrangement shifted, creating internal cavities, with significant changes observed in loop regions.

Conclusions:

  • Cofactor (FMN) removal profoundly influences glycolate oxidase structure, inducing significant flexibility.
  • The apoenzyme adopts a conformation distinct from the holoenzyme, highlighting the role of FMN in stabilizing the active structure.
  • Structural plasticity, particularly in loop regions, is a key feature of glycolate oxidase in its cofactor-depleted state.

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