Open AccessCrystallization and crystal‐packing studies of Chlorella virus deoxyuridine triphosphatase
Author(s) -
Homma Kohei,
Moriyama Hideaki
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109034459
Subject(s) - crystallography , orthorhombic crystal system , histidine , monomer , crystallization , polyethylene glycol , amino acid , chemistry , stereochemistry , crystal structure , biochemistry , polymer , organic chemistry
The 141‐amino‐acid deoxyuridine triphosphatase (dUTPase) from the algal Chlorella virus IL‐3A and its Glu81Ser/Thr84Arg‐mutant derivative Mu‐22 were crystallized using the hanging‐drop vapor‐diffusion method at 298 K with polyethylene glycol as the precipitant. An apo IL‐3A dUTPase with an amino‐terminal T7 epitope tag and a carboxy‐terminal histidine tag yielded cubic P 2 1 3 crystals with unit‐cell parameter a = 106.65 Å. In the presence of dUDP, the enzyme produced thin stacked orthorhombic P 222 crystals with unit‐cell parameters a = 81.0, b = 96.2, c = 132.8 Å. T7‐histidine‐tagged Mu‐22 dUTPase formed thin stacked rectangular crystals. Amino‐terminal histidine‐tagged dUTPases did not crystallize but formed aggregates. Glycyl‐seryl‐tagged dUTPases yielded cubic P 2 1 3 IL‐3A crystals with unit‐cell parameter a = 105.68 Å and hexagonal P 6 3 Mu‐22 crystals with unit‐cell parameters a = 132.07, c = 53.45 Å, γ = 120°. Owing to the Thr84Arg mutation, Mu‐22 dUTPase had different monomer‐to‐monomer interactions to those of IL‐3A dUTPase.