Coupling Computational and Intracellular Screening and Selection Toward Co-compatible cJun and cFos Antagonists
Coupling Computational and Intracellular Screening and Selection Toward Co-compatible cJun and cFos Antagonists
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DOI：
10.1021/acs.biochem.9b00631
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发表时间：
2020-02-04
期刊：
影响因子：
2.9
通讯作者：
Mason, Jody M.
中科院分区：
文献类型：
--
作者：
Lathbridge, Alexander;Michalowska, Anna S.;Mason, Jody M.
碱性亮氨酸拉链（bZIP）蛋白代表了困难的，但令人信服的，致癌的目标，因为许多细胞信号转导级联收敛于他们，在那里他们的功能，以调节特定的基因靶点的转录。bZIPs被广泛认为是包括细胞增殖、凋亡和分化的细胞过程的重要调节剂。一旦这样的经验证的转录调节因子，激活蛋白-1，通常由Fos和Jun家族成员的异源二聚体组成，其中cFos-cJun是最好的描述。它已被证明是许多不同疾病的进展和发展的关键。作为我们的方法的原理证明，我们描述了首次使用一种新的组合的计算机/纤维素肽库筛选平台，该平台有助于衍生出相对于cFos对cJun具有高选择性的序列，同时还避免了同源二聚化。特别地，通过计算筛选了> 6000万个肽，并且根据预测的稳定性对所有潜在的开/关靶标进行排序，从而产生通过细胞内选择进一步细化的尺寸减小的文库。预测衍生的序列与在cJun存在下对cFos具有选择性的第二先前衍生的肽拮抗剂具有有限的串扰。这项研究提供了新的见解，使用多态筛选的能力，联合收割机计算和细胞内的方法，在发展多个兼容的肽，能够满足冲突的设计要求。
Basic leucine-zipper (bZIP) proteins represent difficult, yet compelling, oncogenic targets since numerous cell-signaling cascades converge upon them, where they function to modulate the transcription of specific gene targets. bZIPs are widely recognized as important regulators of cellular processes that include cell proliferation, apoptosis, and differentiation. Once such validated transcriptional regulator, activator protein-1, is typically composed of hetero- dimers of Fos and Jun family members, with cFos-cJun being the best described. It has been shown to be key in the progression and development of a number of different diseases. As a proof-of-principle for our approach, we describe the first use of a novel combined in silico/in cellulo peptide-library screening platform that facilitates the derivation of a sequence that displays high selectivity for cJun relative to cFos, while also avoiding homodimerization. In particular, >60 million peptides were computationally screened and all potential on/off targets ranked according to predicted stability, leading to a reduced size library that was further refined by intracellular selection. The derived sequence is predicted to have limited cross-talk with a second previously derived peptide antagonist that is selective for cFos in the presence of cJun. The study provides new insight into the use of multistate screening with the ability to combine computational and intracellular approaches in evolving multiple cocompatible peptides that are capable of satisfying conflicting design requirements.