Update on the source of phenoloxidase activity in the hemolymph of kuruma prawn Marsupenaeus japonicus.
Update on the source of phenoloxidase activity in the hemolymph of kuruma prawn Marsupenaeus japonicus.
复制标题
日本对虾血淋巴中酚氧化酶活性来源的最新进展。
DOI：
10.1007/s12562-021-01558-x
复制
发表时间：
2021
影响因子：
1.9
通讯作者：
Taro Masuda
中科院分区：
文献类型：
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作者：
辰野竜平;梅枝真人;宮田祐実;出口梨々子;福田翼;古下学;井野靖子;吉川廣幸;高橋洋;長島裕二;Taro Masuda
甲壳动物酚氧化酶（PO）和血蓝蛋白（Hc）被分类为3型铜蛋白。PO催化单酚和二酚化合物的氧化，这是黑化的限速步骤，而HC通常在节肢动物血淋巴中作为双氧转运蛋白。迄今为止，许多研究表明，PO活性在HC中，这是受到它们结构相似性的启发。在这里，PO活性的甲壳类动物的来源，重新审查纯化HC和PO专门从血淋巴的虾类。传统的程序制备节肢动物Hc，其中包括沉淀Hc的超离心和随后的纯化，通过尺寸排阻色谱法，是不能够完全去除血淋巴型PO从Hc。与此相反，用50%饱和度的硫酸铵和随后的疏水色谱分离产生足够纯的HC，其中不含可检测的PO蛋白和几乎没有PO酶活性。这些结果表明，虾类血淋巴中PO活性的主要来源是血淋巴型PO，改进的Hc纯化方法是测定Hc PO活性的较好方法。
Crustacean phenoloxidase (PO) and hemocyanin (Hc) are classified as type 3 copper proteins. PO catalyzes the oxidation of mono- and di-phenol compounds, which is the rate-limiting step of melanization, while Hc generally functions as a dioxygen-transporting protein in the hemolymph of arthropods. To date, many studies have shown PO activity in Hc, which is inspired by their structural similarity. Here, the source of PO activity in crustaceans was re-examined by purifying Hc and PO exclusively from the hemolymph of kuruma prawn. The conventional procedure for the preparation of arthropod Hc, which includes precipitation of Hc by ultracentrifugation and subsequent purification by size exclusion chromatography, was not able to completely remove hemolymph-type PO from Hc. In contrast, fractionation with 50% saturation of ammonium sulfate and subsequent hydrophobic chromatography yielded sufficiently pure Hc, which contained no detectable PO protein and virtually no PO enzymatic activity. These results indicate that the main source of PO activity in the hemolymph of kuruma prawn is hemolymph-type PO and that the improved purification method of Hc is preferable for evaluating the PO activity of Hc.