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Title: Crystallization and Preliminary X-ray Crystallographic Studies of the Oligomeric Death-Domain Complex Between PIDD and RAIDD

Abstract

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 {angstrom} from a native and a Hg-derivative crystal, respectively. The crystals belong to space group P6{sub 5}, with unit-cell parameters a = b = 138.4, c = 207.6 {angstrom}.

Authors:
;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
929889
Report Number(s):
BNL-80471-2008-JA
TRN: US200822%%1071
DOE Contract Number:
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica Section F: Structural Biology and Crystallization Communications; Journal Volume: 63
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; APOPTOSIS; COMPLEXES; CRYSTALLIZATION; CRYSTALS; ESCHERICHIA COLI; INTERACTIONS; LIGHT SCATTERING; PROTEINS; SPACE GROUPS; X-RAY DIFFRACTION; national synchrotron light source

Citation Formats

Park,H., and Wu, H. Crystallization and Preliminary X-ray Crystallographic Studies of the Oligomeric Death-Domain Complex Between PIDD and RAIDD. United States: N. p., 2007. Web. doi:10.1107/S1744309107007889.
Park,H., & Wu, H. Crystallization and Preliminary X-ray Crystallographic Studies of the Oligomeric Death-Domain Complex Between PIDD and RAIDD. United States. doi:10.1107/S1744309107007889.
Park,H., and Wu, H. Mon . "Crystallization and Preliminary X-ray Crystallographic Studies of the Oligomeric Death-Domain Complex Between PIDD and RAIDD". United States. doi:10.1107/S1744309107007889.
@article{osti_929889,
title = {Crystallization and Preliminary X-ray Crystallographic Studies of the Oligomeric Death-Domain Complex Between PIDD and RAIDD},
author = {Park,H. and Wu, H.},
abstractNote = {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 {angstrom} from a native and a Hg-derivative crystal, respectively. The crystals belong to space group P6{sub 5}, with unit-cell parameters a = b = 138.4, c = 207.6 {angstrom}.},
doi = {10.1107/S1744309107007889},
journal = {Acta Crystallographica Section F: Structural Biology and Crystallization Communications},
number = ,
volume = 63,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}