Crystallization and preliminary X-ray crystallographic studies of the oligomeric death-domain complex between PIDD and RAIDD.
Three large macromolecular complexes known as the death-inducing signaling complex (DISC), the apoptosome and the PIDDosome mediate caspase activation in apoptosis signaling pathways. The PIDDosome, which activates caspase-2, is composed of three protein components: PIDD, RAIDD and caspase-2. Within the PIDDosome, the interaction between PIDD and RAIDD is mediated by a homotypic interaction between their death domains (DDs). PIDD DD and RAIDD DD were overexpressed in Escherichia coli with engineered C-terminal His tags. The proteins were purified and mixed to allow complex formation. Gel-filtration and multi-angle light scattering (MALS) analyses showed that the complex is around 150 kDa in solution. The purified PIDD DD-RAIDD DD complex was crystallized at 293 K. X-ray diffraction data were collected to resolutions of 3.2 and 4.0 A from a native and a Hg-derivative crystal, respectively. The crystals belong to space group P6(5), with unit-cell parameters a = b = 138.4, c = 207.6 A.
Park
Hyun Ho
HH
Department of Biochemistry, Weill Medical College and Graduate School of Medical Sciences of Cornell University, New York, NY 10021, USA.
Wu
Hao
H
D017209:Apoptosis; D053420:CRADD Signaling Adaptor Protein / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D053418:Death Domain Receptor Signaling Adaptor Proteins / Q000737:chemistry; D024101:Mitochondrial Proteins / Q000737:chemistry; D020449:Repetitive Sequences, Amino Acid
D053420:CRADD Signaling Adaptor Protein; C104252:CRADD protein, human; D002352:Carrier Proteins; D053418:Death Domain Receptor Signaling Adaptor Proteins; C503369:GRAMD4 protein, human; D024101:Mitochondrial Proteins; C485896:PIDD1 protein, human