Institute of Systems Biology and Bioinformatics, Department of Biomedical Sciences and Engineering, College of Health Sciences and Technology, National Central University, Jhongli 300, Taiwan.
Department of Nutritional Science, Fu Jen Catholic University, New Taipei City 242, Taiwan.
Int J Mol Sci. 2022 Jan 10;23(2):712. doi: 10.3390/ijms23020712.
Cell-penetrating peptides (CPPs) have distinct properties to translocate across cell envelope. The key property of CPPs to translocation with attached molecules has been utilized as vehicles for the delivery of several potential drug candidates that illustrate the significant effect in in-vitro experiment but fail in in-vivo experiment due to selectively permeable nature of cell envelop. Penetratin, a well-known CPP identified from the third α-helix of Antennapedia homeodomain of Drosophila, has been widely used and studied for the delivery of bioactive molecules to treat cancers, stroke, and infections caused by pathogenic organisms. Few studies have demonstrated that penetratin directly possesses antimicrobial activities against bacterial and fungal pathogens; however, the mechanism is unknown. In this study, we have utilized the power of high-throughput proteome microarrays to screen all the potential protein targets of penetratin. proteome microarrays assays of penetratin followed by statistical analysis depicted 123 proteins as the protein targets of penetratin out of ~5800 proteins. To understand the target patterns of penetratin, enrichment analyses were conducted using 123 protein targets. In biological process: ribonucleoprotein complex biogenesis, nucleic acid metabolic process, actin filament-based process, transcription, DNA-templated, and negative regulation of gene expression are a few significantly enriched terms. Cytoplasm, nucleus, and cell-organelles are enriched terms for cellular component. Protein-protein interactions network depicted ribonucleoprotein complex biogenesis, cortical cytoskeleton, and histone binding, which represent the major enriched terms for the 123 protein targets of penetratin. We also compared the protein targets of penetratin and intracellular protein targets of antifungal AMPs (Lfcin B, Histatin-5, and Sub-5). The comparison results showed few unique proteins between penetratin and AMPs. Nucleic acid metabolic process and cellular component disassembly were the common enrichment terms for penetratin and three AMPs. Penetratin shows unique enrichment items that are related to DNA biological process. Moreover, motif enrichment analysis depicted different enriched motifs in the protein targets of penetratin, LfcinB, Histatin-5, and Sub-5.
细胞穿透肽（CPPs）具有独特的特性，可以穿过细胞膜。CPPs 将附着的分子转运到细胞内的关键特性已被用作输送几种潜在药物候选物的载体，这些候选物在体外实验中显示出显著效果，但由于细胞膜的选择性通透性，在体内实验中失败。Penetratin 是一种从果蝇触角同源域的第三α螺旋中鉴定出的著名 CPP，已被广泛用于将生物活性分子递送至治疗癌症、中风和由病原体引起的感染的用途。有一些研究表明 penetratin 直接具有抗细菌和真菌病原体的抗菌活性，但机制尚不清楚。在这项研究中，我们利用高通量蛋白质组微阵列的力量筛选 penetratin 的所有潜在蛋白质靶标。对 penetratin 进行蛋白质组微阵列测定，然后进行统计分析，描绘出 penetratin 的 123 种蛋白质作为蛋白质靶标，而约 5800 种蛋白质中只有 123 种。为了了解 penetratin 的靶标模式，使用 123 种蛋白质靶标进行了富集分析。在生物过程中：核糖核蛋白复合物的生物发生、核酸代谢过程、肌动蛋白丝为基础的过程、转录、DNA 模板和基因表达的负调节是几个显著富集的术语。细胞质、细胞核和细胞器官是细胞成分的富集术语。蛋白质-蛋白质相互作用网络描绘了核糖核蛋白复合物的生物发生、皮质细胞骨架和组蛋白结合，这代表了 penetratin 的 123 种蛋白质靶标的主要富集术语。我们还比较了 penetratin 的蛋白质靶标和抗真菌 AMPs（Lfcin B、Histatin-5 和 Sub-5）的细胞内蛋白质靶标。比较结果表明 penetratin 和 AMPs 之间有几个独特的蛋白质。核酸代谢过程和细胞成分解体是 penetratin 和三种 AMPs 的共同富集术语。Penetratin 显示出与 DNA 生物过程相关的独特富集项目。此外，基序富集分析描绘了 penetratin、LfcinB、Histatin-5 和 Sub-5 的蛋白质靶标中的不同富集基序。
