Protein Kinase C (PKC) is a member of the AGC subfamily of kinases and regulates a wide array of signaling pathways and physiological processes. Protein–protein interactions involving PKC and its scaffolding partners dictate the spatiotemporal dynamics of PKC activity, including its access to activating second messenger molecules and potential substrates. While the A Kinase Anchoring Protein (AKAP) family of scaffold proteins universally bind PKA, several were also found to scaffold PKC, thereby serving to tune its catalytic output. Targeting these scaffolding interactions can further shed light on the effect of subcellular compartmentalization on PKC signaling. Here we report the development of two hydrocarbon stapled peptides, CSTAD5 and CSTAD6, that are cell permeable and bind PKC to disrupt PKC–gravin complex formation in cells. Both constrained peptides downregulate PMA-induced cytoskeletal remodeling that is mediated by the PKC–gravin complex as measured by cell rounding. Further, these peptides downregulate PKC substrate phosphorylation and cell motility. To the best of our knowledge, no PKC-selective AKAP disruptors have previously been reported and thus CSTAD5 and CSTAD6 are novel disruptors of PKC scaffolding by AKAPs and may serve as powerful tools for dissecting AKAP-localized PKC signaling.

中文翻译：

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AKAP 信号复合物靶向破坏 PKC

蛋白激酶 C (PKC) 是激酶 AGC 亚家族的成员，可调节多种信号通路和生理过程。涉及 PKC 及其支架伙伴的蛋白质-蛋白质相互作用决定了 PKC 活性的时空动态，包括其对激活第二信使分子和潜在底物的访问。虽然支架蛋白的 A 激酶锚定蛋白 (AKAP) 家族普遍结合 PKA，但也发现有几个支架蛋白可以支架 PKC，从而调节其催化输出。针对这些支架相互作用可以进一步阐明亚细胞区室化对 PKC 信号传导的影响。在这里，我们报告了两种烃钉肽 CSTAD5 和 CSTAD6 的开发，它们具有细胞渗透性并结合 PKC 以破坏 PKC –gravin 复合物在细胞中的形成。两种受约束的肽都下调 PMA 诱导的由 PKC - gravin 复合物介导的细胞骨架重塑，如通过细胞舍入测量。此外，这些肽下调 PKC 底物磷酸化和细胞运动。据我们所知，以前没有报道过 PKC 选择性 AKAP 干扰物，因此 CSTAD5 和 CSTAD6 是 AKAP 对 PKC 支架的新型干扰物，可以作为剖析 AKAP 定位的 PKC 信号传导的有力工具。