气候变化领域集成服务门户(CCPortal): Discovery of a caspase cleavage motif antibody reveals insights into noncanonical inflammasome function

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DOI	10.1073/pnas.2018024118
	Discovery of a caspase cleavage motif antibody reveals insights into noncanonical inflammasome function
	Davies C.W.; Stowe I.; Phung Q.T.; Ho H.; Bakalarski C.E.; Gupta A.; Zhang Y.; Lill J.R.; Payandeh J.; Kayagaki N.; Koerber J.T.
发表日期	2021
ISSN	00278424
卷号	118 期号:12
英文摘要	Inflammasomes sense a number of pathogen and host damage signals to initiate a signaling cascade that triggers inflammatory cell death, termed pyroptosis. The inflammatory caspases (1/4/5/ 11) are the key effectors of this process through cleavage and activation of the pore-forming protein gasdermin D. Caspase-1 also activates proinflammatory interleukins, IL-1β and IL-18, via proteolysis. However, compared to the well-studied apoptotic caspases, the identity of substrates and therefore biological functions of the inflammatory caspases remain limited. Here, we construct, validate, and apply an antibody toolset for direct detection of neoC termini generated by inflammatory caspase proteolysis. By combining rabbit immune phage display with a set of degenerate and defined target peptides, we discovered two monoclonal antibodies that bind peptides with a similar degenerate recognition motif as the inflammatory caspases without recognizing the canonical apoptotic caspase recognition motif. Crystal structure anal-yses revealed the molecular basis of this strong yet paradoxical degenerate mode of peptide recognition. One antibody selectively immunoprecipitated cleaved forms of known and unknown inflammatory caspase substrates, allowing the identification of over 300 putative substrates of the caspase-4 noncanonical inflammasome, including caspase-7. This dataset will provide a path toward developing blood-based biomarkers of inflammasome activation. Overall, our study establishes tools to discover and detect inflammatory caspase substrates and functions, provides a workflow for designing antibody reagents to study cell signaling, and extends the growing evidence of biological cross talk between the apoptotic and inflammatory caspases. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Antibody engineering \| noncanonical inflammasome \| inflammatory caspase
语种	英语
scopus关键词	caspase; caspase 7; inflammasome; interleukin 18; interleukin 1beta; monoclonal antibody; Article; carboxy terminal sequence; cell assay; controlled study; crystal structure; enzyme activation; enzyme specificity; enzyme substrate; immunoprecipitation; intracellular signaling; mass spectrometry; molecular recognition; nonhuman; phage display; priority journal; protein cleavage; protein degradation; protein function; protein motif; spliceosome
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180151
作者单位	Department of Antibody Engineering, Genentech, Inc., South San Francisco, CA 94080, United States; Department of Physiological Chemistry, Genentech, Inc., South San Francisco, CA 94080, United States; Department of Microchemistry Proteomics and Lipidomics, Genentech, Inc., South San Francisco, CA 94080, United States; Department of Structural Biology, Genentech, Inc., South San Francisco, CA 94080, United States; Department of Early Discovery Biochemistry, Genentech, Inc., South San Francisco, CA 94080, United States
推荐引用方式 GB/T 7714	Davies C.W.,Stowe I.,Phung Q.T.,et al. Discovery of a caspase cleavage motif antibody reveals insights into noncanonical inflammasome function[J],2021,118(12).
APA	Davies C.W..,Stowe I..,Phung Q.T..,Ho H..,Bakalarski C.E..,...&Koerber J.T..(2021).Discovery of a caspase cleavage motif antibody reveals insights into noncanonical inflammasome function.Proceedings of the National Academy of Sciences of the United States of America,118(12).
MLA	Davies C.W.,et al."Discovery of a caspase cleavage motif antibody reveals insights into noncanonical inflammasome function".Proceedings of the National Academy of Sciences of the United States of America 118.12(2021).