Purification, crystallization and preliminary X-ray crystallographic study of the L-fuculose-1-phosphate aldolase (FucA) from Thermus thermophilus HB8.
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.
Jeyakanthan
Jeyaraman
J
Biometal Science Laboratory, RIKEN Harima Institute, 1-1-1 Kouto, Sayo-gun, Hyogo 679-5148, Japan. kanthan@spring8.or.jp
Taka
Junichiro
J
Kikuchi
Akihiro
A
Kuroishi
Chizu
C
Yutani
Katsuhide
K
Shiro
Yoshitugu
Y
D000446:Aldehyde-Lyases / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004099:Dihydroxyacetone Phosphate / Q000737:chemistry; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry*; D008027:Light; D011487:Protein Conformation; D012542:Scattering, Radiation; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D004099:Dihydroxyacetone Phosphate; D000446:Aldehyde-Lyases; C103622:FucA protein, bacteria; D005634:Fructose-Bisphosphate Aldolase; C036027:L-fuculosephosphate aldolase