Expression, purification, crystallization of fragments from the C-terminal region of DFF45/ICAD.
DFF45/ICAD, which forms a heterodimer with DFF40/CAD as its DNase inhibitor and chaperone, plays a key role in nuclei DNA fragmentation in apoptosis. Several fragments from the C-terminal region of DFF45/ICAD have been cloned and expressed in Escherichia coli as His-tagged proteins. After purification to homogeneity, the recombinant proteins of three fragments were crystallized by the hanging-drop vapor-diffusion method. Of these, a crystal of DFF45c1 diffracted to 3.4 A in a capillary at 277 K and crystals of DFF45c2 diffracted to 3.2 A at cryotemperature using synchrotron radiation.
Ding
Haitao
H
Life Science College, Peking University, Beijing 100871, People's Republic of China.
Qiu
Shihong
S
Li
Songlin
S
Symersky
Jindrich
J
Green
Todd J
TJ
Luo
Ming
M
D000595:Amino Acid Sequence; D051017:Apoptosis Regulatory Proteins; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction / Q000379:methods
D051017:Apoptosis Regulatory Proteins; D010446:Peptide Fragments; D011506:Proteins; C109664:caspase-activated DNase inhibitor