Open AccessCrystallization and preliminary X‐ray crystallographic studies of Drep‐3, a DFF‐related protein from Drosophila melanogaster
Author(s) -
Park Hyun Ho,
Tookes Hansel Emory,
Wu Hao
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106018422
Subject(s) - homotetramer , dna fragmentation , drosophila melanogaster , crystallography , crystallization , recombinant dna , apoptosis , microbiology and biotechnology , biology , schneider 2 cells , chemistry , biochemistry , programmed cell death , gene , protein subunit , rna , rna interference , organic chemistry
During apoptosis, DNA fragmentation is mainly mediated by the caspase‐activated DFF40 nuclease. DFF40 exists as a heterodimeric complex with its inhibitor DFF45. Upon apoptosis induction, DFF45 is cleaved by caspases to allow DFF40 activation. Drep‐3 is a recently identified regulator of the DFF40 system in Drosophila melanogaster . Here, Drep‐3 was expressed with a C‐terminal His tag in Escherichia coli and the protein was purified to homogeneity. Multi‐angle light‐scattering analysis showed that Drep‐3 is a homotetramer in solution. Native and selenomethionine‐substituted Drep‐3 proteins were crystallized at 293 K and X‐ray diffraction data were collected to 2.8 and 3.0 Å resolution, respectively. The crystals belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 56.9, b = 125.4, c = 168.7 Å. The asymmetric unit is estimated to contain one homotetramer.