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Self-oligomerization of ASC PYD Domain Prevents the Assembly of Inflammasome In Vitro

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Abstract

NALP3 inflammasome, which is an inflammatory caspase-activating complex, is composed of three proteins: NALP3 (an NOD-like receptor), an apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and caspase-1. NALP3 senses danger signals, while ASC is an adaptor molecule containing two protein interaction modules: pyrin domain (PYD) and caspase recruitment domain (CARD). Caspase-1 is a cysteine protease that uses cysteine as a nucleophile and has a CARD domain for protein interaction. During inflammasome formation, the ASC adaptor acts as a bridge between caspase and NOD-like receptor (NLR) by offering the CARD for CARD–CARD interactions and PYD for PYD–PYD interactions. In the current study, we successfully purified and characterized NALP3 PYD and ASC PYD. The results showed that ASC PYD easily self-oligomerized under physiological conditions, and this self-oligomerization of the ASC PYD prevented complex formation with NALP3 PYD in vitro.

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Acknowledgments

This study was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) of the Ministry of Education, Science and Technology (2013009083) and a grant from the Korea Healthcare Technology R&D project, Ministry of Health & Welfare, Republic of Korea (HI13C1449).

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  1. Department of Biochemistry, School of Biotechnology and Graduate School of Biochemistry, Yeungnam University, Gyeongsan, South Korea

    Kannan Badri Narayanan, Tae-Ho Jang & Hyun Ho Park

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  1. Kannan Badri Narayanan
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  2. Tae-Ho Jang
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  3. Hyun Ho Park
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Correspondence to Hyun Ho Park.

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Kannan Badri Narayanan and Tae-Ho Jang contributed equally to this work.

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Narayanan, K.B., Jang, TH. & Park, H.H. Self-oligomerization of ASC PYD Domain Prevents the Assembly of Inflammasome In Vitro. Appl Biochem Biotechnol 172, 3902–3912 (2014). https://doi.org/10.1007/s12010-014-0819-0

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