Journal
JOURNAL OF BIOTECHNOLOGY
Volume 151, Issue 4, Pages 335-342Publisher
ELSEVIER
DOI: 10.1016/j.jbiotec.2011.01.003
Keywords
Apoptosis; Inflammation; Death domain superfamily; Purification; Refolding
Categories
Funding
- Ministry for Health, Welfare and Family Affairs, Republic of Korea [A100325]
- Korea Health Promotion Institute [A100325] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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The death domain (DD) superfamily comprising the death domain (DD) subfamily, the death effector domain (DED) subfamily, the caspase recruitment domain (CARD) subfamily and the pyrin domains (PYD) subfamily is one of the largest classes of protein interaction modules and plays a pivotal role in the apoptosis, inflammation, and immune cell signaling pathways. Despite the biological importance of the death domain superfamily, structural and in vitro biochemical studies have been limited because these domains are prone to aggregate under physiological conditions. Here, we describe a generalized method, termed semi-refolding. that is particularly applicable for purification of the functional death domain superfamily. The recombinant proteins Caspase-1 CARD, AIM2 PYD, NALP3 PYD, and RIM DD from inclusion bodies were successfully purified using this method. (C) 2011 Elsevier B.V. All rights reserved.
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