Crystallization and X-ray analysis of D-threonine aldolase from Chlamydomonas reinhardtii

Acta Crystallogr F Struct Biol Commun. 2017 Feb 1;73(Pt 2):86-89. doi: 10.1107/S2053230X1602063X. Epub 2017 Jan 19.

Abstract

D-Threonine aldolase from the green alga Chlamydomonas reinhardtii (CrDTA) catalyzes the interconversion of several β-hydroxy-D-amino acids (e.g. D-threonine) and glycine plus the corresponding aldehydes. Recombinant CrDTA was overexpressed in Escherichia coli and purified to homogeneity; it was subsequently crystallized using the hanging-drop vapour-diffusion method at 295 K. Data were collected and processed at 1.85 Å resolution. Analysis of the diffraction pattern showed that the crystal belonged to space group P1, with unit-cell parameters a = 64.79, b = 74.10, c = 89.94 Å, α = 77.07, β = 69.34, γ = 71.93°. The asymmetric unit contained four molecules of CrDTA. The Matthews coefficient was calculated to be 2.12 Å3 Da-1 and the solvent content was 41.9%.

Keywords: Chlamydomonas reinhardtii; crystallization; d-amino acids; d-threonine aldolase.

MeSH terms

  • Algal Proteins / chemistry*
  • Algal Proteins / genetics
  • Algal Proteins / metabolism
  • Amino Acid Sequence
  • Chlamydomonas reinhardtii / chemistry*
  • Chlamydomonas reinhardtii / enzymology
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Glycine Hydroxymethyltransferase / chemistry*
  • Glycine Hydroxymethyltransferase / genetics
  • Glycine Hydroxymethyltransferase / metabolism
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • X-Ray Diffraction

Substances

  • Algal Proteins
  • Recombinant Proteins
  • Glycine Hydroxymethyltransferase