Feldman Michal, Levy Dan
The Shraga Segal Department of Microbiology, Immunology and Genetics, National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Be'er-Sheva 84105, Israel.
National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Be'er-Sheva 84105, Israel.
Oncotarget. 2017 Dec 21;9(4):4875-4885. doi: 10.18632/oncotarget.23591. eCollection 2018 Jan 12.
A large body of evidence accumulating in the past few years indicates the physiological significance of non-histone proteins lysine methylation, catalyzed by protein lysine methyl transferases (PKMTs). Dysregulation of these enzymes was shown to contribute to the development and progression of numerous diseases. SETD6 lysine methylatransferase was recently shown to participate in essential cellular processes, such as the NFkB pathway, oxidative stress and also the Wnt signaling cascade. In order to test the effect of blocking SETD6 catalytic activity, we used the peptide inhibition method, which is considered highly specific and can potentially target almost any protein. We designed a 15 amino acids peptide based on the sequence of the RelA protein (residues 302-316), containing the lysine that is methylated by SETD6. To enable cellular intake, the designed peptide was fused to a cell penetrating peptide (CPP) vp22. The vp22-RelA302-316 peptide showed direct and specific interaction with SETD6 . This interaction was shown to inhibit SETD6 methyltransferase activity. SETD6 catalytic blockage by the peptide was also observed in cells upon treatment with the vp22-RelA302-316, resulting in induced cellular migration and proliferation. This new insight into the activity of a methylation inhibitory peptide could represent a milestone in the development of therapeutic tools, which can be of use in physiological cases where administration of cell proliferation is required.
在过去几年中积累的大量证据表明，由蛋白质赖氨酸甲基转移酶（PKMTs）催化的非组蛋白赖氨酸甲基化具有生理意义。这些酶的失调被证明与多种疾病的发生和发展有关。SETD6赖氨酸甲基转移酶最近被证明参与重要的细胞过程，如NFkB途径、氧化应激以及Wnt信号级联反应。为了测试阻断SETD6催化活性的效果，我们使用了肽抑制方法，该方法被认为具有高度特异性，并且几乎可以潜在地靶向任何蛋白质。我们基于RelA蛋白的序列（残基302 - 316）设计了一种15个氨基酸的肽，其中包含被SETD6甲基化的赖氨酸。为了实现细胞摄取，将设计的肽与细胞穿透肽（CPP）vp22融合。vp22 - RelA302 - 316肽显示出与SETD6直接且特异性相互作用。这种相互作用被证明可抑制SETD6甲基转移酶活性。在用vp22 - RelA302 - 316处理细胞后，也观察到该肽对SETD6的催化阻断作用，导致细胞迁移和增殖增加。对甲基化抑制肽活性的这一新见解可能代表了治疗工具开发中的一个里程碑，可用于需要促进细胞增殖的生理情况。
