OBJECTIVE To design protein phosphatase 1 (PP1)-disrupting peptides covalently coupled to inert cell-penetrating peptides (CPPs) as sychnologically organized bioportide constructs as a strategy to modulate sperm motility. DESIGN Experimental study. SETTING Academic research laboratory. PATIENT(S)/ANIMAL(S) Normozoospermic men providing samples for routine analysis and Holstein Frisian bulls. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Effect of the bioportides on the activity and interactions of PP1γ2-a PP1 isoform expressed exclusively in testicular germ cells and sperm-and on sperm vitality and motility. RESULT(S) PP1-disrupting peptides were designed based on the sequences from: 1) a sperm-specific PP1 interactor (A kinase anchor protein 4); and 2) a PP1 inhibitor (protein phosphatase inhibitor 2). Those sequences were covalently coupled to inert CPPs as bioportide constructs, which were successfully delivered to the flagellum of sperm cells to induce a marked impact on PP1γ2 activity and sperm motility. Molecular modeling studies further facilitated the identification of an optimized PP1-binding sequence and enabled the development of a modified stop-sperm bioportide with reduced size and increased potency of action. In addition, a bioportide mimetic of the unique 22-amino acid C-terminus of PP1γ2 accumulated within spermatozoa to significantly reduce sperm motility and further define the PP1γ2-specific interactome. CONCLUSION(S) These investigations demonstrate the utility of CPPs to deliver peptide sequences that target unique protein-protein interactions in spermatozoa to achieve a significant impact upon spermatozoa motility, a key prognostic indicator of male fertility.

中文翻译：

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使用 bioportides 作为靶向精子活力的新方法破坏蛋白磷酸酶 1 复合物

目的 设计蛋白磷酸酶 1 (PP1) 破坏肽与惰性细胞穿透肽 (CPP) 共价偶联，作为按顺序组织的生物蛋白结构，作为调节精子活力的策略。设计 实验研究。设置学术研究实验室。PATIENT(S)/ANIMAL(S) 正常精子症男性和荷斯坦弗里斯兰公牛提供用于常规分析的样本。干预措施 无。主要结果测量（S） bioportides 对 PP1γ2-a PP1 同种型的活性和相互作用的影响，仅在睾丸生殖细胞和精子中表达 - 以及对精子活力和活力的影响。结果 基于以下序列设计了 PP1 破坏肽：1) 精子特异性 PP1 相互作用子（激酶锚蛋白 4）；和 2) PP1 抑制剂（蛋白磷酸酶抑制剂 2）。这些序列与惰性 CPP 共价偶联，作为生物端口构建体，成功递送至精子细胞的鞭毛，对 PP1γ2 活性和精子活力产生显着影响。分子建模研究进一步促进了优化的 PP1 结合序列的鉴定，并能够开发出尺寸更小、作用更强的改良停止精子生物转运体。此外，在精子中积累的 PP1γ2 独特的 22 个氨基酸的 C 端的 bioportide 模拟物可显着降低精子活力并进一步定义 PP1γ2 特异性相互作用组。结论（S）这些研究证明了 CPP 可用于传递靶向精子中独特蛋白质-蛋白质相互作用的肽序列，从而对精子运动产生显着影响，