Profiling human protein degradome delineates cellular responses to proteasomal inhibition and reveals a feedback mechanism in regulating proteasome homeostasis
Profiling human protein degradome delineates cellular responses to proteasomal inhibition and reveals a feedback mechanism in regulating proteasome homeostasis
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DOI：
10.1038/cr.2014.122
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发表时间：
2014-09
期刊：
影响因子：
44.1
通讯作者：
Tao Yu;Yonghui Tao;Meiqiang Yang;Peng Chen;Xiaobo Gao;Yanbo Zhang;Tao Zhang;Zi-Jun Chen;Jian Hou;Yan Zhang;Kangcheng Ruan;Hongyan Wang;Ronggui Hu
中科院分区：
文献类型：
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作者：
Tao Yu;Yonghui Tao;Meiqiang Yang;Peng Chen;Xiaobo Gao;Yanbo Zhang;Tao Zhang;Zi-Jun Chen;Jian Hou;Yan Zhang;Kangcheng Ruan;Hongyan Wang;Ronggui Hu
蛋白质周转的整体变化（蛋白质降解组）构成了细胞对内在或外在刺激反应的核心部分。然而，分析蛋白质降解产物仍然具有技术挑战性。最近，蛋白酶体的抑制，例如，通过使用硼替佐米（BTZ），已成为治疗多发性骨髓瘤和其他人类恶性肿瘤的主要化疗策略，但BTZ药物作用和肿瘤耐药机制的系统理解尚未实现。在这里，我们开发并应用了一种基于双荧光的蛋白质周转试验（ProTA），以定量分析BTZ诱导的蛋白酶体抑制后人类蛋白质降解的全球变化。ProTA和随后的网络分析描绘了BTZ化疗中BTZ作用和肿瘤耐药性的潜在分子基础。最后，BTZ与靶向ProTA鉴定的BTZ作用中的关键基因或途径的药物的联合使用降低了多发性骨髓瘤细胞中的BTZ抗性。值得注意的是，BTZ稳定蛋白酶体亚基PSMC 1和蛋白酶体组装因子PSMD 10，这表明以前未被充分认识的调节蛋白酶体稳态的机制。因此，ProTA是一种用于分析人类蛋白质降解组的新型工具，以阐明药物作用和耐药性的潜在机制，这可能有助于开发针对蛋白质稳态的治疗方法来治疗人类疾病。
Global change in protein turnover (protein degradome) constitutes a central part of cellular responses to intrinsic or extrinsic stimuli. However, profiling protein degradome remains technically challenging. Recently, inhibition of the proteasome, eg, by using bortezomib (BTZ), has emerged as a major chemotherapeutic strategy for treating multiple myeloma and other human malignancies, but systematic understanding of the mechanisms for BTZ drug action and tumor drug resistance is yet to be achieved. Here we developed and applied a dual-fluorescence-based Protein Turnover Assay (ProTA) to quantitatively profile global changes in human protein degradome upon BTZ-induced proteasomal inhibition. ProTA and subsequent network analyses delineate potential molecular basis for BTZ action and tumor drug resistance in BTZ chemotherapy. Finally, combined use of BTZ with drugs targeting the ProTA-identified key genes or pathways in BTZ action reduced BTZ resistance in multiple myeloma cells. Remarkably, BTZ stabilizes proteasome subunit PSMC1 and proteasome assembly factor PSMD10, suggesting a previously under-appreciated mechanism for regulating proteasome homeostasis. Therefore, ProTA is a novel tool for profiling human protein degradome to elucidate potential mechanisms of drug action and resistance, which might facilitate therapeutic development targeting proteostasis to treat human disorders.