title: Generalized semi-refolding methods for purification of the functional death domain superfamily.
abstract: 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 RIP1 DD from inclusion bodies were successfully purified using this method.
authors.affiliation: School of Biotechnology and Graduate School of Biochemistry, Yeungnam University, Gyeongsan, Republic of Korea.
authors.forename: Tae-ho
authors.initials: TH
authors.lastname: Jang
authors.forename: Hyun Ho
authors.initials: HH
authors.lastname: Park
mesh_terms: D002352:Carrier Proteins / Q000302:isolation & purification; D020170:Caspase 1 / Q000302:isolation & purification; D002942:Circular Dichroism / Q000379:methods; D004268:DNA-Binding Proteins; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D028861:Nuclear Pore Complex Proteins / Q000302:isolation & purification; D009687:Nuclear Proteins / Q000302:isolation & purification; D058849:Protein Refolding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000302:isolation & purification*; D016601:RNA-Binding Proteins / Q000302:isolation & purification
chemical_list: C119724:AGFG1 protein, human; C410167:AIM2 protein, human; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D011506:Proteins; D016601:RNA-Binding Proteins; D020170:Caspase 1