, Volume 15, Issue 12, pp 1444–1452 | Cite as

In vitro reconstitution of the interactions in the PIDDosome

  • Tae-ho Jang
  • Chao Zheng
  • Hao Wu
  • Ju-Hong Jeon
  • Hyun Ho ParkEmail author
Original Paper


Caspase-2 is critical for genotoxic stress induced apoptosis and is activated by formation of the PIDDosome, an oligomeric caspase-2 activating complex. The PIDDosome comprises three protein components, PIDD, RAIDD, and caspase-2. RAIDD contains both a death domain (DD) and a caspase recruitment domain (CARD). It acts as the bridge to recruit PIDD using the DD: DD interaction and to recruit caspase-2 via the CARD: CARD interaction. Here we report biochemical characterization and in vitro reconstitution of the core interactions in the PIDDosome. We show that RAIDD CARD and RAIDD DD interact with their binding partners, caspase-2 CARD and PIDD DD, respectively. However, full-length RAIDD fails to interact with either caspase-2 CARD or PIDD DD under a physiological buffer condition. We reveal that this lack of interaction of full-length RAIDD is not due to its tendency to aggregate under the physiological buffer condition, as decreasing full-length RAIDD aggregation using high salt or high pH is not able to promote complex formation. Instead, full-length RAIDD forms both binary and ternary complexes under a low salt condition. Successful in vitro reconstitution of the ternary complex provides a basis for further structural studies of the PIDDosome.


Apoptosis Inflammation Caspase-2 RAIDD PIDD PIDDosome 



This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) of the Ministry of Education, Science and Technology (2010-0015311 to HHP) and by the National Institute of Health (RO1 AI076927 to HW).


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Tae-ho Jang
    • 1
  • Chao Zheng
    • 2
  • Hao Wu
    • 2
  • Ju-Hong Jeon
    • 3
  • Hyun Ho Park
    • 1
    Email author
  1. 1.Department of BiochemistrySchool of Biotechnology and Graduate School of Biochemistry at Yeungnam UniversityGyeongsanSouth Korea
  2. 2.Department of BiochemistryWeill Medical College and Graduate School of Medical Sciences of Cornell UniversityNew YorkUSA
  3. 3.Department of PhysiologySeoul National University College of MedicineSeoulSouth Korea

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