Department of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Science and Technology, Wyb. Wyspianskiego 27, 50-370, Wroclaw, Poland.
NCI-designated Cancer Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, 92037, USA.
Sci Rep. 2018 Oct 30;8(1):15998. doi: 10.1038/s41598-018-34476-7.
Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) belongs to the CD clan of cysteine proteases. MALT1 is a unique enzyme among this clan because it recognizes the basic amino acid arginine in the P1 pocket. Previous studies carried out with natural amino acids revealed the substrate specificity of the P4-P1 pockets of MALT1 but have provided only limited information about the catalytic preferences of this enzyme. In this study, we exploited Hybrid Combinatorial Substrate Library and Internally Quenched Fluorescence substrate technologies to interrogate the extended substrate specificity profile of the S5-S2' active site pockets using unnatural amino acids. This strategy resulted in the design of a peptide-based fluorogenic substrate, which exhibited significant activity toward MALT1. Subsequently, the substrate sequence was further utilized to develop potent, irreversible activity-based probes.
黏膜相关淋巴组织淋巴瘤易位蛋白 1（MALT1）属于半胱氨酸蛋白酶 CD 家族。MALT1 是该家族中一种独特的酶，因为它能识别 P1 口袋中的碱性氨基酸精氨酸。先前使用天然氨基酸进行的研究揭示了 MALT1 的 P4-P1 口袋的底物特异性，但仅提供了有关该酶催化偏好的有限信息。在这项研究中，我们利用混合组合底物文库和内部猝灭荧光底物技术，使用非天然氨基酸来探究 S5-S2'活性位点口袋的扩展底物特异性谱。该策略导致设计了一种基于肽的荧光底物，该底物对 MALT1 表现出显著的活性。随后，进一步利用该底物序列来开发有效的、不可逆的基于活性的探针。
