Computational and biochemical studies of isothiocyanates as inhibitors of proteasomal cysteine deubiquitinases in human cancer cells.
Computational and biochemical studies of isothiocyanates as inhibitors of proteasomal cysteine deubiquitinases in human cancer cells.
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DOI：
10.1002/jcb.27157
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发表时间：
2018-11
影响因子：
4
通讯作者：
Dou QP
中科院分区：
文献类型：
--
作者：
Ahmed ZSO;Li X;Li F;Cheaito HA;Patel K;Mosallam EM;Elbargeesy GAEH;Dou QP
异硫氰酸酯（ITCs）是一种天然的化学保护物质，广泛存在于十字花科蔬菜中。然而，ITCs的癌症相关靶点和分子机制仍不清楚。我们假设，ITCs，亲电，可以与催化三联体（CYS，HIS和ASP）的蛋白酶体半胱氨酸去泛素化酶USP 14和UCHL 5，最终抑制其活动。在目前的研究中，我们利用这种可能性，使用人类乳腺癌和前列腺癌细胞模型进行计算对接和生化验证试验。对接结果表明异硫氰酸苄酯（BITC）、异硫氰酸苯乙酯（PEITC）和DL-萝卜硫素（SFN）是比USP 14更有效的UCHL 5抑制剂，并且这些ITC可以与UCHL 5和USP 14的催化三联体相互作用。事实上，Ub-VS测定证实了每个ITC对UCHL 5和USP 14的泛素结合活性的抑制活性。我们还发现ITC对USP-14和UCHL 5活性的抑制导致USP 14和UCHL 5蛋白水平升高，但第三种19 S-DUB，RPN 11没有，这表明反馈环激活，并进一步支持ITC是蛋白酶体半胱氨酸DUB的抑制剂。ITCs抑制DUB的同时，泛素化蛋白表达显著增加，并诱导细胞凋亡、抑制细胞增殖和抑制细胞侵袭。我们的研究结果ITCs作为蛋白酶体半胱氨酸DUB抑制剂应该提供有见地的信息，设计，发现和开发有效的，具体的19 S-DUB抑制剂的癌症治疗。
Isothiocyanates (ITCs) are natural chemoprotective products found abundantly in cruciferous vegetables. However, the cancer-relevant targets and molecular mechanisms of ITCs remain unclear. We hypothesize that ITCs, as electrophiles, can interact with the catalytic triads (CYS, HIS and ASP) of the proteasomal cysteine deubiquitinases USP14 and UCHL5, ultimately inhibiting their activities. In the current study, we exploited this possibility by performing both computational docking and biochemical validation assays using human breast and prostate cancer cell models. Docking results suggest that benzyl isothiocyanate (BITC), phenethyl isothiocyanate (PEITC) and DL-Sulforaphane (SFN) are more potent inhibitors of UCHL5 than USP14, and these ITCs could interact with the catalytic triads of UCHL5 and USP14. Indeed, Ub-VS assay confirmed the inhibitory activity of each ITC on the ubiquitin-binding activity of UCHL5 and USP14. We also found that inhibition of USP-14 and UCHL5 activities by the ITCs caused increased levels of USP14 and UCHL5 proteins, but not the third 19S-DUB, RPN11, suggesting feedback loop activation and further supporting that ITCs are inhibitors of proteasomal cysteine DUBs. Associated with DUB inhibition by ITCs, ubiquitinated proteins were significantly increased, accompanied with induction of apoptosis, inhibition of proliferation and suppression of cell invasion. Our findings of ITCs as proteasomal cysteine DUB inhibitors should provide insightful information for designing, discovering and developing potent, specific 19S-DUB inhibitors for cancer therapies.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.3
作者：
Aressy, Bernadette;Jullien, Denis;Ducommun, Bernard
通讯作者：
Ducommun, Bernard
DOI：
10.1111/j.1742-4658.2011.08393.x
发表时间：
2011-12
期刊：
The FEBS journal
影响因子：
--
作者：
Maiti TK;Permaul M;Boudreaux DA;Mahanic C;Mauney S;Das C
通讯作者：
Das C
影响因子：
64.8
作者：
Lander, Gabriel C.;Estrin, Eric;Matyskiela, Mary E.;Bashore, Charlene;Nogales, Eva;Martin, Andreas
通讯作者：
Martin, Andreas
影响因子：
5.2
作者：
Gupta, Parul;Kim, Bonglee;Kim, Sung-Hoon;Srivastava, Sanjay K.
通讯作者：
Srivastava, Sanjay K.