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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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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Herausgeber
May, M.J.*
Schlagwörter
MALT1; Cysteine protease; Activity assay; Autoimmunity; Inflammation; Lymphoma
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Konferenztitel
NF-kappa B: Methods and Protocols
Zeitschrift
Methods in Molecular Biology
Quellenangaben
Band: 1280,
Seiten: 239-246
Verlag
Springer
Verlagsort
Berlin [u.a.]
Begutachtungsstatus
Peer reviewed
Institut(e)
Research Unit Signaling and Translation (SAT)
Institute of Molecular Toxicology and Pharmacology (TOX)
Institute of Molecular Toxicology and Pharmacology (TOX)