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Selection of cDNA candidates that induce oligomerization of NLRP3 using a chimeric receptor approach

Paper ID Volume ID Publish Year Pages File Format Full-Text
20257 43165 2015 8 PDF Available
Title
Selection of cDNA candidates that induce oligomerization of NLRP3 using a chimeric receptor approach
Abstract

Since diverse cellular events are regulated by protein oligomerization, identification of molecules that affect oligomerization of a target protein is important for understanding cellular physiology and developing therapeutics. In this study, we aimed to screen cDNA candidates that induce oligomerization of NLRP3, which is one of the important inflammatory sensor proteins, in mammalian cytoplasm. In our screening method, the chimera composed of NLRP3 and the kinase domain of c-kit, one of the receptor tyrosine kinases (RTKs) activated by oligomerization, is expressed in cytoplasm of an IL-3-dependent mammalian cell line. The cells are then transduced with a cDNA library, and cultured in the absence of IL-3. If the transduced cDNA is a NLRP3 activator, the kinase domain of the NLRP3-c-kit chimera is activated by oligomerization, which induces cell growth even in the absence of IL-3. Using this system, constitutive oligomers of two NLRP3 variants were clearly detected by cell growth. Furthermore, cDNA screening resulted in identification of three distinct cDNAs that are potential candidates of NLRP3 activators. These results demonstrate the utility of our chimeric receptor-based system for screening candidates that induce oligomerization of a target protein.

Keywords
Protein oligomer; Chimeric receptor; Tyrosine phosphorylation; cDNA library; Mammalian cell
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Selection of cDNA candidates that induce oligomerization of NLRP3 using a chimeric receptor approach
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 120, Issue 2, August 2015, Pages 223–230
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us