Published: Vol 3, Iss 14, Jul 20, 2013 DOI: 10.21769/BioProtoc.821 Views: 12038
Reviewed by: Lin FangFanglian He
Protocol Collections
Comprehensive collections of detailed, peer-reviewed protocols focusing on specific topics
Related protocols
Measuring Protein Synthesis during Cell Cycle by Azidohomoalanine (AHA) Labeling and Flow Cytometric Analysis
Koshi Imami and Tomoharu Yasuda
Apr 20, 2019 8673 Views
Analysis of the Ubiquitination and Phosphorylation of Vangl Proteins
Di Feng [...] Bo Gao
Oct 20, 2022 2426 Views
Isoform-specific, Semi-quantitative Determination of Highly Homologous Protein Levels via CRISPR-Cas9-mediated HiBiT Tagging
Kristina Seiler [...] Mario P. Tschan
Jul 20, 2023 1376 Views
Abstract
MALT1(Mucosa associated lymphoid tissue protein 1) is an important adapter protein for the NF-kB driven lymphocyte activation and the development and survival of distinct B-cell lymphoma entities. In addition MALT1 is a cysteine protease that structurally resembles caspases while having a different substrate preference and mechanism of activation. This paracaspase activity of MALT1 has been shown to be critical for an optimal NF-kB activation and survival of the aggressive ABC-DLBCL (Activated B cell-type of diffuse large B cell lymphoma), which highlights the protease as an attractive therapeutic target for the treatment of distinct B-cell lymphomas and immune diseases like rheumatoid arthritis or multiple sclerosis. In this protocol we describe a fluorogenic cleavage assay, which can be used to measure endogenous and also ectopic MALT1 activity. To this end, cellular MALT1 needs to be precipitated from the lysed cells via antibody immunoprecipitation and subsequently incubated with a fluorogenic substrate peptide. The MALT1 cleavage assay has been developed to directly determine the activity profile of MALT1 in the course of the adaptive immune response as well as in pathological signaling in lymphoid malignancies. In addition, the MALT1 activity assay has been successfully used to monitor cellular MALT1 inhibition with small molecule inhibitors.
Keywords: ProteaseMaterials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol was adapted from two previous publications (Kloo et al., 2011; Nagel et al., 2012). The work was supported by a grant of the ‘Deutsche Krebshilfe e.V.’ to DK.
References
Article Information
Copyright
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Nagel, D. and Krappmann, D. (2013). Measurement of Endogenous MALT1 Activity. Bio-protocol 3(14): e821. DOI: 10.21769/BioProtoc.821.
Category
Cancer Biology > General technique > Biochemical assays
Biochemistry > Protein > Activity
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Tips for asking effective questions
+ Description
Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.
Share
Bluesky
X
Copy link