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Company: EMD Millipore

Catalog#: 407950

Isoform-specific, Semi-quantitative Determination of Highly Homologous Protein Levels via CRISPR-Cas9-mediated HiBiT Tagging

Kristina Seiler, Sreoshee Rafiq and Mario P. Tschan

Published online: 2023-07-20

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Measurement of Endogenous MALT1 Activity

Author:

Daniel Nagel and Daniel Krappmann,

Date:

2013-07-20

[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 ... [摘要] MALT1（粘膜相关淋巴组织蛋白1）是NF-kB驱动的淋巴细胞活化和不同B细胞淋巴瘤实体的发展和存活的重要衔接蛋白。此外，MALT1是半胱氨酸蛋白酶，其在结构上类似于半胱天冬酶，同时具有不同的底物偏好和激活机制。已显示MALT1的这种半胱氨酸蛋白酶活性对于侵袭性ABC-DLBCL（活化的B细胞型弥漫性大B细胞淋巴瘤）的最佳NF-kB活化和存活是至关重要的，其突出显示蛋白酶作为有吸引力的治疗靶标治疗不同的B细胞淋巴瘤和免疫性疾病如类风湿性关节炎或多发性硬化。在该协议中，我们描述了荧光切割测定，其可以用于测量内源性以及异位MALT1活性。为此，细胞MALT1需要通过抗体免疫沉淀从裂解的细胞中沉淀，随后与荧光底物肽孵育。已经开发了MALT1切割测定以直接测定MALT1在适应性免疫应答过程中以及淋巴恶性肿瘤中的病理信号传导中的活性谱。此外，MALT1活性测定已经成功地用于用小分子抑制剂监测细胞MALT1抑制。

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