A Set of Activity-Based Probes to Visualize Human (Immuno)proteasome Activities
A Set of Activity-Based Probes to Visualize Human (Immuno)proteasome Activities
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DOI：
10.1002/anie.201509092
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发表时间：
2016-03-18
影响因子：
16.6
通讯作者：
Overkleeft, Herman S.
中科院分区：
文献类型：
--
作者：
de Bruin, Gerjan;Xin, Bo Tao;Overkleeft, Herman S.
蛋白酶体是各种癌症和自身免疫性疾病的治疗靶点。组成型表达的蛋白酶体有三个活性位点，1c，2c和5c。类肉瘤组织也表达免疫蛋白酶体亚单位1i、2i和5i。快速和同时测量这些催化亚基的活性将有助于发现新的抑制剂，改善临床试验中蛋白酶体抑制剂的分析，并简化亚基表达的分析。在这项工作中，我们提出了一个鸡尾酒的活性为基础的探针，使同时基于凝胶的检测所有六个催化人类蛋白酶体亚基。我们使用这种混合物来开发1c，2c，5c和2i的特异性抑制剂，比较临床蛋白酶体抑制剂的活性位点特异性，并证明许多血液恶性肿瘤主要表达免疫蛋白酶体。此外，我们表明，选择性和完全抑制5i和1i是细胞毒性的急性淋巴细胞白血病（ALL）患者的原代细胞。
Proteasomes are therapeutic targets for various cancers and autoimmune diseases. Constitutively expressed proteasomes have three active sites, 1c, 2c, and 5c. Lymphoid tissues also express the immunoproteasome subunits 1i, 2i, and 5i. Rapid and simultaneous measurement of the activity of these catalytic subunits would assist in the discovery of new inhibitors, improve analysis of proteasome inhibitors in clinical trials, and simplify analysis of subunit expression. In this work, we present a cocktail of activity-based probes that enables simultaneous gel-based detection of all six catalytic human proteasome subunits. We used this cocktail to develop specific inhibitors for 1c, 2c, 5c, and 2i, to compare the active-site specificity of clinical proteasome inhibitors, and to demonstrate that many hematologic malignancies predominantly express immunoproteasomes. Furthermore, we show that selective and complete inhibition of 5i and 1i is cytotoxic to primary cells from acute lymphocytic leukemia (ALL) patients.