Elucidation of the crystal structure of Coriolopsis caperata laccase: restoration of the structure and activity of the native enzyme from the T2-depleted form by copper ions

Acta Crystallogr D Biol Crystallogr. 2015 Apr;71(Pt 4):854-61. doi: 10.1107/S1399004715001595. Epub 2015 Mar 26.

Abstract

Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. A new laccase was isolated from the basidiomycete Coriolopsis caperata strain 0677 and its amino-acid sequence was determined. According to its physicochemical properties and spectroscopic features, the laccase from C. caperata is a high redox-potential blue laccase. Attempts to crystallize the native enzyme were unsuccessful. The copper type 2-depleted (T2D) laccase was prepared and crystallized. The structure of T2D laccase from C. caperata was solved at 1.6 Å resolution, and attempts to reconstruct the T2 copper centre were performed using Cu(+) and Cu(2+) ions. The structure of T2D+Cu(+) laccase was solved at 1.89 Å resolution. It was shown that the T2D+Cu(+) laccase structure contained four copper ions in the active site. Reconstruction could not be achieved when the T2D laccase crystals were treated with CuSO4.

Keywords: Coriolopsis caperata; copper type 2-depleted laccase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalytic Domain
  • Copper / chemistry*
  • Copper / metabolism
  • Coriolaceae / chemistry
  • Coriolaceae / enzymology*
  • Crystallography, X-Ray
  • Laccase / chemistry*
  • Laccase / metabolism
  • Models, Molecular
  • Oxidation-Reduction
  • Protein Conformation

Substances

  • Copper
  • Laccase

Associated data

  • PDB/4JHU
  • PDB/4JHV