title: Activity-based probes for detection of active MALT1 paracaspase in immune cells and lymphomas.
abstract: MALT1 paracaspase is activated upon antigen receptor stimulation to promote lymphocyte activation. In addition, deregulated MALT1 protease activity drives survival of distinct lymphomas such as the activated B cell type of diffuse large B cell lymphoma (ABC-DLBCL). Here, we designed fluorophore or biotin-coupled activity based-probes (ABP) that covalently modify the active center of MALT1. MALT1-ABPs are exclusively labeling an active modified full length form of MALT1 upon T cell stimulation. Further, despite the CARMA1 requirement for initial MALT1 activation, the MALT1-ABPs show that protease activity is not confined to the high-molecular CARMA1-BCL10-MALT1 (CBM) complex. Using biotin-coupled ABPs, we developed a robust assay for sensitive and selective detection of active MALT1 in cell lines, primary lymphocytes, and DLBCL tumor biopsies. Taken together, MALT1-ABPs represent powerful chemical tools to measure cellular MALT1 activation, determine efficacy of small molecule inhibitors, and classify lymphomas based on MALT1 activity status.
authors.affiliation: Research Unit Cellular Signal Integration, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstaedter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Eitelhuber
authors.affiliation: Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Vosyka
authors.affiliation: Research Unit Cellular Signal Integration, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstaedter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Nagel
authors.affiliation: Research Unit Cellular Signal Integration, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstaedter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Miriam
authors.initials: M
authors.lastname: Bognar
authors.affiliation: Department of Pathology, Charité - University Medicine Berlin, Hindenburgdamm 30, 12200 Berlin, Germany.
authors.forename: Dido
authors.initials: D
authors.lastname: Lenze
authors.affiliation: Department of Biochemistry, Gene Center, Ludwig-Maximilian University, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Lammens
authors.affiliation: Department of Biochemistry, Gene Center, Ludwig-Maximilian University, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Schlauderer
authors.affiliation: Research Unit Cellular Signal Integration, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstaedter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Hlahla
authors.affiliation: Department of Biochemistry, Gene Center, Ludwig-Maximilian University, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
authors.forename: Karl-Peter
authors.initials: KP
authors.lastname: Hopfner
authors.affiliation: Department of Medicine A, Translational Oncology, Albert-Schweitzer Campus 1, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Lenz
authors.affiliation: Department of Pathology, Charité - University Medicine Berlin, Hindenburgdamm 30, 12200 Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hummel
authors.affiliation: Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany.
authors.forename: Steven H L
authors.initials: SH
authors.lastname: Verhelst
authors.affiliation: Research Unit Cellular Signal Integration, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstaedter Landstraße 1, 85764 Neuherberg, Germany. Electronic address: daniel.krappmann@helmholtz-muenchen.de.
authors.forename: Daniel
authors.initials: D
authors.lastname: Krappmann
mesh_terms: D001710:Biotin / Q000737:chemistry; D015153:Blotting, Western; D053468:CARD Signaling Adaptor Proteins / Q000378:metabolism; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D057930:Click Chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006162:Guanylate Cyclase / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D016403:Lymphoma, Large B-Cell, Diffuse / Q000201:enzymology* / Q000473:pathology*; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D000074884:Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry; D013601:T-Lymphocytes / Q000166:cytology / Q000201:enzymology* / Q000276:immunology
chemical_list: D053468:CARD Signaling Adaptor Proteins; D015335:Molecular Probes; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D012235:Rhodamines; C005358:tetramethylrhodamine; D001710:Biotin; D020169:Caspases; C400109:MALT1 protein, human; D000074884:Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein; C426242:CARD11 protein, human; D006162:Guanylate Cyclase