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

doi:10.1107/S1744309107007889

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

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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:: Park,H.; Wu, H.

Publication Date:: Mon Jan 01 00:00:00 EST 2007

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.

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                    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.

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                    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.

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@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}

}

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DOI: 10.1107/S1744309107007889

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