A Multipronged Approach Establishes Covalent Modification of β-Tubulin as the Mode of Action of Benzamide Anti-cancer Toxins.
A Multipronged Approach Establishes Covalent Modification of β-Tubulin as the Mode of Action of Benzamide Anti-cancer Toxins.
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DOI：
10.1021/acs.jmedchem.0c01482
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发表时间：
2020-11-25
影响因子：
7.3
通讯作者：
McFadden DG
中科院分区：
文献类型：
--
作者：
Povedano JM;Rallabandi R;Bai X;Ye X;Liou J;Chen H;Kim J;Xie Y;Posner B;Rice L;De Brabander JK;McFadden DG
表型高通量筛选鉴定出一种具有抗小细胞肺癌细胞活性的苯甲酰胺小分子。设计了一种“可点击”的苯甲酰胺探针，它不可逆地结合单个50 kDa的细胞蛋白，经质谱仪鉴定为β-微管蛋白。此外，一系列苯甲酰胺类似物的抗癌效力与探针竞争密切相关，表明β-微管蛋白是功能靶点。更多的证据表明，苯甲酰胺在秋水仙素结合部位共价修饰了Cys239。与这一机制一致，苯甲酰胺损害了含有Cys239的β-微管蛋白形成的微管的生长，但对编码Ser239的β3微管蛋白没有影响。因此，我们设计了一种含醛类似物，能够将Ser239捕获到β3微管蛋白中，推测为半缩醛。使用正向遗传学策略，我们鉴定了具有β-微管蛋白Thr238Ala突变的苯甲酰胺耐药细胞系，该突变足以诱导复合耐药。公开的化学探针有助于识别其他秋水仙素结合部位，这是结构多样化小分子的常见目标。
A phenotypic high-throughput screen identified a benzamide small molecule with activity against small cell lung cancer cells. A “clickable” benzamide probe was designed that irreversibly bound a single 50 kDa cellular protein, identified by mass spectrometry as β-tubulin. Moreover, the anti-cancer potency of a series of benzamide analogs strongly correlated with probe competition, indicating that β-tubulin was the functional target. Additional evidence suggested that benzamides covalently modified Cys239 within the colchicine binding site. Consistent with this mechanism, benzamides impaired growth of microtubules formed with β-tubulin harboring Cys239, but not β3 tubulin encoding Ser239. We therefore designed an aldehyde-containing analog capable of trapping Ser239 in β3 tubulin, presumably as a hemiacetal. Using a forward genetics strategy, we identified benzamide-resistant cell lines harboring a Thr238Ala mutation in β-tubulin sufficient to induce compound resistance. The disclosed chemical probes are useful to identify other colchicine site binders, a frequent target of structurally diverse small molecules.
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影响因子：
64.5
作者：
McFadden DG;Papagiannakopoulos T;Taylor-Weiner A;Stewart C;Carter SL;Cibulskis K;Bhutkar A;McKenna A;Dooley A;Vernon A;Sougnez C;Malstrom S;Heimann M;Park J;Chen F;Farago AF;Dayton T;Shefler E;Gabriel S;Getz G;Jacks T
通讯作者：
Jacks T
影响因子：
3.3
作者：
MACHIN, NA;LEE, JM;BARNES, G
通讯作者：
BARNES, G
一个孤立的扣子TOG域抑制微管灾难并促进救援。
影响因子：
3.3
作者：
Majumdar S;Kim T;Chen Z;Munyoki S;Tso SC;Brautigam CA;Rice LM
通讯作者：
Rice LM
影响因子：
16
作者：
Jost M;Chen Y;Gilbert LA;Horlbeck MA;Krenning L;Menchon G;Rai A;Cho MY;Stern JJ;Prota AE;Kampmann M;Akhmanova A;Steinmetz MO;Tanenbaum ME;Weissman JS
通讯作者：
Weissman JS
影响因子：
7.7
作者：
Geyer EA;Burns A;Lalonde BA;Ye X;Piedra FA;Huffaker TC;Rice LM
通讯作者：
Rice LM