Purification, crystallization and preliminary X-ray crystallographic study of the L-fuculose-1-phosphate aldolase (FucA) from Thermus thermophilus HB8

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Dec 1;61(Pt 12):1075-7. doi: 10.1107/S1744309105036766. Epub 2005 Nov 24.

Abstract

Fuculose phosphate aldolase catalyzes the reversible cleavage of L-fuculose-1-phosphate to dihydroxyacetone phosphate and L-lactaldehyde. The protein from Thermus thermophilus HB8 is a biological tetramer with a subunit molecular weight of 21 591 Da. Purified FucA has been crystallized using sitting-drop vapour-diffusion and microbatch techniques at 293 K. The crystals belong to space group P4, with unit-cell parameters a = b = 100.94, c = 45.87 A. The presence of a dimer of the enzyme in the asymmetric unit was estimated to give a Matthews coefficient (VM) of 2.7 A3 Da(-1) and a solvent content of 54.2%(v/v). Three-wavelength diffraction MAD data were collected to 2.3 A from zinc-containing crystals. Native diffraction data to 1.9 A resolution have been collected using synchrotron radiation at SPring-8.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde-Lyases / chemistry
  • Bacterial Proteins / chemistry*
  • Crystallization
  • Crystallography, X-Ray / methods*
  • Dihydroxyacetone Phosphate / chemistry
  • Fructose-Bisphosphate Aldolase / chemistry*
  • Light
  • Protein Conformation
  • Scattering, Radiation
  • Temperature
  • Thermus thermophilus / enzymology*
  • X-Ray Diffraction

Substances

  • Bacterial Proteins
  • Dihydroxyacetone Phosphate
  • Aldehyde-Lyases
  • FucA protein, bacteria
  • Fructose-Bisphosphate Aldolase
  • L-fuculosephosphate aldolase