Open Access Green as soon as Postprint is submitted to ZB.
Detection of recombinant and cellular MALT1 paracaspase activity.
Methods Mol. Biol. 1280, 239-246 (2015)
MALT1 (mucosa-associated lymphoid tissue protein 1) is a key regulator of antigen-induced NF-κB activation in the adaptive immune response. Activation of proteolytic activity of the MALT1 paracaspase was shown to boost the immune response. Additionally, MALT1 proteolytic activity is essential for the survival of MALT1-dependent lymphoma, such as the activated B-cell type (ABC) of diffuse large B-cell lymphoma (DLBCL) or MALT lymphoma. The functional impact of MALT1 paracaspase on T-cell activation and lymphomagenesis suggests that MALT1 is a promising therapeutic target for the treatment of autoimmune diseases and distinct lymphoma entities. To evaluate the requirement of MALT1 in further detail, direct measurement of its activity status is of great importance. We have established a fluorogenic cleavage assay which can be used to measure activity of recombinant and cellular MALT1. Here we describe the basis of the cleavage assay and include a detailed protocol for recombinant production of MALT1 and also the cellular immunoprecipitation of endogenous MALT1 to determine its proteolytic activity.
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Publication type
Article: Journal article
Document type
Scientific Article
Editors
May, M.J.*
Keywords
MALT1; Cysteine protease; Activity assay; Autoimmunity; Inflammation; Lymphoma
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Conference Title
NF-kappa B: Methods and Protocols
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 1280,
Pages: 239-246
Publisher
Springer
Publishing Place
Berlin [u.a.]
Reviewing status
Peer reviewed
Institute(s)
Research Unit Signaling and Translation (SAT)
Institute of Molecular Toxicology and Pharmacology (TOX)
Institute of Molecular Toxicology and Pharmacology (TOX)