Open AccessPurification, crystallization and preliminary X‐ray crystallographic study of the l ‐fuculose‐1‐phosphate aldolase (FucA) from Thermus thermophilus HB8
Author(s) -
Jeyakanthan Jeyaraman,
Taka Junichiro,
Kikuchi Akihiro,
Kuroishi Chizu,
Yutani Katsuhide,
Shiro Yoshitugu
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105036766
Subject(s) - thermus thermophilus , crystallography , crystallization , dimer , aldolase a , tetramer , chemistry , thermus , dihydroxyacetone phosphate , x ray crystallography , phosphate , diffraction , enzyme , thermophile , escherichia coli , organic chemistry , biochemistry , physics , optics , gene
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 P 4, with unit‐cell parameters a = b = 100.94, c = 45.87 Å. The presence of a dimer of the enzyme in the asymmetric unit was estimated to give a Matthews coefficient ( V M ) of 2.7 Å 3 Da −1 and a solvent content of 54.2%( v / v ). Three‐wavelength diffraction MAD data were collected to 2.3 Å from zinc‐containing crystals. Native diffraction data to 1.9 Å resolution have been collected using synchrotron radiation at SPring‐8.