定量化学蛋白质组学揭示了异硫氰酸苯乙酯可共价作用于 BID,从而促进细胞凋亡。

IF 6.1 2区 生物学 Q1 CELL BIOLOGY
Xiaoshu Dong, Xinqian Yu, Minghao Lu, Yaxin Xu, Liyan Zhou, Tao Peng
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引用次数: 0

摘要

十字花科蔬菜中天然存在的异硫氰酸盐(ITCs),如异硫氰酸苄酯(BITC)、异硫氰酸苯乙酯(PEITC)和莱菔硫烷(SFN),因其在体外和体内具有良好的抗癌活性而引起了研究人员的极大兴趣。虽然诱导细胞凋亡被认为在 ITCs 的抗癌作用中起着关键作用,但 ITC 诱导细胞凋亡的特定蛋白靶点和相关上游事件仍不为人知。在本研究中,我们提出了一套从 BITC、PEITC 和 SFN 提取并配备生物正交炔柄的化学探针,用于系统分析活癌细胞中的 ITC 靶蛋白。利用基于竞争的定量化学蛋白质组学方法,我们确定了一系列富含凋亡等生物过程的 ITC 候选靶蛋白。我们发现 Bcl-2 家族的凋亡调节因子 BID 的 N 端半胱氨酸被 ITC 共价修饰。功能特性分析表明,PEITC 与 BID N 端半胱氨酸的共价结合会导致蛋白质构象的改变,并破坏 BID N 端和 C 端之间的自我抑制作用,从而释放高活性的 C 端片段,发挥下游的促凋亡效应。同样,PEITC 可促进 BID 的裂解和线粒体转运,从而强烈诱导细胞凋亡。我们进一步发现,N-端半胱氨酸的突变会损害 BID 的 N- 端和 C- 端相互作用,从而解除自我抑制并增强其凋亡活性。总之,我们的化学蛋白质组学分析和功能研究不仅揭示了 BID 是 PEITC 诱导细胞凋亡上游事件的主要靶点,而且还发现了一种新的分子机制,即 BID 第一螺旋内的 N 端半胱氨酸参与调控其促凋亡潜能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Quantitative chemical proteomics reveals that phenethyl isothiocyanate covalently targets BID to promote apoptosis.

Naturally occurring isothiocyanates (ITCs) found in cruciferous vegetables, such as benzyl isothiocyanate (BITC), phenethyl isothiocyanate (PEITC), and sulforaphane (SFN), have attracted significant research interest for their promising anti-cancer activity in vitro and in vivo. While the induction of apoptosis is recognized to play a key role in the anti-cancer effects of ITCs, the specific protein targets and associated upstream events underlying ITC-induced apoptosis remain unknown. In this study, we present a set of chemical probes that are derived from BITC, PEITC, and SFN and equipped with bioorthogonal alkynyl handles to systematically profile the target proteins of ITCs in live cancer cells. Using a competition-based quantitative chemical proteomics approach, we identify a range of candidate target proteins of ITCs enriched in biological processes such as apoptosis. We show that BID, an apoptosis regulator of the Bcl-2 family, is covalently modified by ITCs on its N-terminal cysteines. Functional characterization demonstrates that covalent binding to N-terminal cysteines of BID by PEITC results in conformational changes of the protein and disruption of the self-inhibitory interaction between N- and C-terminal regions of BID, thus unleashing the highly active C-terminal segment to exert downstream pro-apoptotic effects. Consistently, PEITC promotes the cleavage and mitochondrial translocation of BID, leading to a strong induction of apoptosis. We further show that mutation of N-terminal cysteines impairs the N- and C-terminal interaction of BID, relieving the self-inhibition and enhancing its apoptotic activity. Overall, our chemical proteomics profiling and functional studies not only reveal BID as the principal target of PEITC in mediating upstream events for the induction of apoptosis, but also uncover a novel molecular mechanism involving N-terminal cysteines within the first helix of BID in regulating its pro-apoptotic potential.

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来源期刊
Cell Death Discovery
Cell Death Discovery Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
8.30
自引率
1.40%
发文量
468
审稿时长
9 weeks
期刊介绍: Cell Death Discovery is a multidisciplinary, international, online-only, open access journal, dedicated to publishing research at the intersection of medicine with biochemistry, pharmacology, immunology, cell biology and cell death, provided it is scientifically sound. The unrestricted access to research findings in Cell Death Discovery will foster a dynamic and highly productive dialogue between basic scientists and clinicians, as well as researchers in industry with a focus on cancer, neurobiology and inflammation research. As an official journal of the Cell Death Differentiation Association (ADMC), Cell Death Discovery will build upon the success of Cell Death & Differentiation and Cell Death & Disease in publishing important peer-reviewed original research, timely reviews and editorial commentary. Cell Death Discovery is committed to increasing the reproducibility of research. To this end, in conjunction with its sister journals Cell Death & Differentiation and Cell Death & Disease, Cell Death Discovery provides a unique forum for scientists as well as clinicians and members of the pharmaceutical and biotechnical industry. It is committed to the rapid publication of high quality original papers that relate to these subjects, together with topical, usually solicited, reviews, editorial correspondence and occasional commentaries on controversial and scientifically informative issues.
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