The effect of helix-inducing constraints and downsizing upon a transcription block survival-derived functional cJun antagonist.
The effect of helix-inducing constraints and downsizing upon a transcription block survival-derived functional cJun antagonist.
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DOI：
10.1016/j.xcrp.2022.101077
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发表时间：
2022-10-19
期刊：
影响因子：
--
通讯作者：
Mason JM
中科院分区：
文献类型：
--
作者：
Brennan A;Leech JT;Kad NM;Mason JM
cJun的抑制被确立为一种有前景的治疗方法，特别是在癌症中。我们最近开发了“转录阻断存活”（TBS）筛选平台，通过消除其与同源DNA结合的能力来获得转录因子活性的功能性肽拮抗剂。使用TBS，我们筛选了> 131，000个成员的肽文库，以选择结合cJun并阻止12-O-十四酰基佛波醇-13-乙酸酯响应元件（TRE）DNA结合的63-mer肽。接下来将迭代截短与侧链环化的系统探索相结合，以获得最小活性序列。所得的双内酰胺化序列小>40%，并保留低nM靶亲和力（平衡结合常数[KD] = 0.2 vs 9.7 nM），在功能性拮抗作用所需的酸性区域具有8个残基。然而，即使是适度的C-末端截短也会导致功能丧失。该肽在功能上拮抗cJun（半数最大抑制浓度[IC 50] = 13 vs 45 μM），并且相对于其非内酰胺化对应物和HingeW，在人血清中的稳定性显著更高。先前衍生的cJun肽拮抗剂已被优化HW 1的系统性缩小决定了最小活性序列添加螺旋诱导内酰胺桥以恢复因缩小HW 30而损失的功效保留活性，尺寸减小，并显示出增加的血清稳定性Brennan et al.报道了通过去除>40%的残基并添加螺旋诱导侧链内酰胺桥来优化经验证的致癌转录调节因子cJun的转录阻断存活测定衍生的肽拮抗剂。优化的肽保留了功能并显示出增加的血清稳定性，为未来的应用产生了更像药物的分子。
Inhibition of cJun is established as a promising therapeutic approach, particularly in cancer. We recently developed the “transcription block survival” (TBS) screening platform to derive functional peptide antagonists of transcription factor activity by ablating their ability to bind to cognate DNA. Using TBS, we screened a >131,000-member peptide library to select a 63-mer peptide that bound cJun and prevented 12-O-tetradecanoylphorbol-13-acetate response element (TRE) DNA binding. Iterative truncation was next combined with a systematic exploration of side-chain cyclization to derive a minimal active sequence. The resulting dual lactamized sequence was >40% smaller and retained low nM target affinity (equilibrium binding constant [KD] = 0.2 versus 9.7 nM), with 8 residues at the acidic region required for functional antagonism. However, even modest C-terminal truncation resulted in functional loss. The peptide functionally antagonizes cJun (half-maximal inhibitory concentration [IC50] = 13 versus 45 μM) and is considerably more stable in human serum relative to its non-lactamized counterpart and HingeW. A previously derived cJun peptide antagonist has been optimized Systematic downsizing of HW1 determines the minimal active sequence Helix-inducing lactam bridges are added to restore efficacy lost from downsizing HW30 retains activity, is reduced in size, and displays increased serum stability Brennan et al. report that a transcription block survival assay-derived peptide antagonist of validated oncogenic transcriptional regulator cJun is optimized by removing >40% of residues and adding helix-inducing side-chain lactam bridges. The optimized peptide retains function and displays increased serum stability, producing a more drug-like molecule for future applications.
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影响因子：
14.8
作者：
Baker EG;Bartlett GJ;Crump MP;Sessions RB;Linden N;Faul CF;Woolfson DN
通讯作者：
Woolfson DN
影响因子：
3.7
作者：
Rao T;Ruiz-Gómez G;Hill TA;Hoang HN;Fairlie DP;Mason JM
通讯作者：
Mason JM
影响因子：
8
作者：
Brennan, Andrew;Leech, James T;Kad, Neil M;Mason, Jody M
通讯作者：
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影响因子：
4.8
作者：
Olive, M;Williams, SC;Vinson, C
通讯作者：
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影响因子：
18.3
作者：
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通讯作者：
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