Prophenoloxidase is a metalloprotinase and closely associated with the copper proteins family like hemocyanin; both are extracellular proteins that are structurally similar and functionally vary in their roles. In this study, we used the mud crab Scylla serrata as a model organism to explore the mechanism of phenoloxidase (PO) and structure analysis through in silico approach also attempting to discover the chemical inhibitors for the protein molecule from different databases. We have purified metalloprotein, prophenoloxidase (proPO), exhibits the 76 kDa molecular weight in SDS-PAGE, widely distributed in hemocytes and consisted of 2 homologous polypeptide chains, proPO1 and proPO2. Enhanced PO activity was observed when using microbial pathogen-associated molecular patterns (MAMPs) and metals as an activator. In vitro and in silico approaches were employed to characterize the structural and functional properties of phenoloxidases, and also we used the virtually screened ligands to check the activity of the protein. Through virtual screening method to screen the new ligands to bind with proPO from zinc and binding databases, the binding interactions are also analyzed via docking scores. This present report paves the way to develop the new immunomodulatory ligands targeting toward the prophenoloxidase and also gives new insights about the structure of the protein.

中文翻译：

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锯缘青蟹中酚氧化酶原的表征和结构分析及通过虚拟筛选发现新型化学抑制剂

酚氧化酶原是一种金属蛋白酶，与血蓝蛋白等铜蛋白家族密切相关；两者都是结构相似且功能不同的细胞外蛋白。在这项研究中，我们以泥蟹 Scylla serrata 作为模式生物，通过计算机方法探索酚氧化酶 (PO) 的机制和结构分析，并试图从不同的数据库中发现蛋白质分子的化学抑制剂。我们纯化了金属蛋白，酚氧化酶原 (proPO)，在 SDS-PAGE 中的分子量为 76 kDa，广泛分布于血细胞中，由 2 条同源多肽链 proPO1 和 proPO2 组成。当使用微生物病原体相关分子模式 (MAMP) 和金属作为活化剂时，观察到增强的 PO 活性。体外和计算机中的方法被用来表征酚氧化酶的结构和功能特性，我们还使用虚拟筛选的配体来检查蛋白质的活性。通过虚拟筛选方法从锌和结合数据库中筛选与 proPO 结合的新配体，还通过对接评分分析结合相互作用。本报告为开发针对酚氧化酶原的新免疫调节配体铺平了道路，并提供了有关蛋白质结构的新见解。结合相互作用也通过对接分数进行分析。本报告为开发针对酚氧化酶原的新免疫调节配体铺平了道路，并提供了有关蛋白质结构的新见解。结合相互作用也通过对接分数进行分析。本报告为开发针对酚氧化酶原的新免疫调节配体铺平了道路，并提供了有关蛋白质结构的新见解。