Graduate Institute of Systems Biology and Bioinformatics, and Department of Biomedical Sciences and Engineering, College of Health Sciences and Technology, National Central University, Jhongli 32001, Taiwan.
Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
Int J Mol Sci. 2021 Jan 13;22(2):760. doi: 10.3390/ijms22020760.
Antimicrobial peptides (AMPs) are intensively studied in terms of alternative drugs. Sub5 is a synthetic 12-mer AMP with substitutions of five amino acids of bactenecin 2A (Bac2A), a linear-ized bactenecin variant of bovine. Sub5 is highly effective against fungi with an ability to trans-locate cell membrane, but its targets are unknown. Systematic analysis of Sub5 targets will facil-itate our understanding on its mechanism of action. In this study, we used high-throughput proteome microarrays to explore the potential protein targets of Sub5. The screening results showed 128 potential protein targets of Sub5. Bioinformatics analysis of protein targets of Sub5 revealed significant gene ontology (GO) enrichment in actin related pro-cess of "actin filament-based process", "actin filament organization", "actin cortical patch or-ganization", regulation of "actin filament bundle assembly". Moreover, the other enriched cat-egories in GO enrichment mostly contained actin associate proteins. In total, 11 actin-associated proteins were identified in the protein targets of Sub5. Protein family (PFAM) enrichment anal-ysis shows protein domain enriched in actin binding, i.e., "Cytoskeletal-regulatory complex EF hand (helix E-loop-helix F motif)". Being consistent with GO analysis, Search Tool for the Re-trieval of Interacting Genes/Proteins (STRING) analysis of the protein targets of Sub5 showed ac-tin network with involvement of 15 protein targets. Along with actin-network, STRING analysis showed protein-protein interaction network in ribonucleoprotein, transcription and translation, chromosome, histone, and ubiquitin related, DNA repair, and chaperone. Multiple Expression motifs for Motif Elicitation (MEME) suite provided a consensus binding motif of [ED][ED]EEE[ED][ED][ED][ED][ED], in total of 75 protein targets of Sub5. This motif was present in 9 out of 15 actin-related proteins identified among protein targets of Sub5.
抗菌肽 (AMPs) 作为替代药物受到了广泛的研究。Sub5 是一种合成的 12 肽 AMP，其取代了牛源杀菌肽 2A (Bac2A) 的 5 个氨基酸。Sub5 对真菌具有高度的有效性，能够穿透细胞膜，但它的靶点尚不清楚。对 Sub5 靶点的系统分析将有助于我们理解其作用机制。在本研究中，我们使用高通量蛋白质组微阵列来探索 Sub5 的潜在蛋白质靶点。筛选结果显示 Sub5 有 128 个潜在的蛋白质靶点。Sub5 蛋白质靶点的生物信息学分析显示，在“肌动蛋白丝为基础的过程”、“肌动蛋白丝组织”、“肌动蛋白皮质斑组织”和“肌动蛋白丝束组装的调节”的“肌动蛋白相关过程”中，GO 显著富集。此外，GO 富集的其他丰富类别大多包含肌动蛋白相关蛋白。共有 11 种肌动蛋白相关蛋白被鉴定为 Sub5 的蛋白质靶点。蛋白质家族 (PFAM) 富集分析显示富含肌动蛋白结合的蛋白结构域，即“细胞骨架调节复合物 EF 手 (螺旋 E-环-螺旋 F 基序)”。与 GO 分析一致，Sub5 蛋白质靶点的 Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) 分析显示，肌动蛋白网络涉及 15 个蛋白质靶点。除肌动蛋白网络外，STRING 分析还显示了与核糖核蛋白、转录和翻译、染色体、组蛋白和泛素相关、DNA 修复和伴侣相关的蛋白质-蛋白质相互作用网络。Motif Elicitation (MEME) suite 提供的多个表达基序提供了一个共识结合基序 [ED][ED]EEE[ED][ED][ED][ED][ED]，共有 75 个 Sub5 的蛋白质靶点。该基序存在于 Sub5 蛋白质靶点中鉴定的 15 种肌动蛋白相关蛋白中的 9 种。
