University of Maryland Center for Environmental Science, Institute of Marine and Environmental Technology, 701 East Pratt Street, Columbus Center, Suite 236, Baltimore, MD 21202, USA.
Fish Shellfish Immunol. 2012 Jan;32(1):16-25. doi: 10.1016/j.fsi.2011.08.023. Epub 2011 Oct 28.
The full-length cDNA (1362 nucleotides, GenBank JF736621) encoding an extracellular copper zinc superoxide dismutase initially isolated from an EST library of the blue crab Callinectes sapidus was characterized using 3' RACE and named Cas-ecCuZnSOD-2. The open reading frame of Cas-ecCuZnSOD-2 contains 203 deduced amino acids with the conserved active catalytic center for copper and zinc binding and the post-translational modification at two putative N-glycosylation and nine phosphorylation sites. Overall, the deduced amino acids of Cas-ecCuZnSOD-2 shared only 35% sequence identity with that of Cas-ecCuZnSOD (GenBank AF264031) which was previously found in C. sapidus, while it showed ∼75% sequence identity to Scylla paramamosain ecCuZnSOD (GenBank FJ774661). The expression profile of Cas-ecCuZnSOD-2 and the other three C. sapidus SODs: ecCuZn, cytMn- and mitMn SODs was largely ubiquitous among the tested tissues obtained from a juvenile female at intermolt: brain, eyestalk ganglia, pericardial organs, and thoracic ganglia complex (nervous system); hepatopancreas (digestive system); heart, artery and hemocytes (circulatory system); gill and antennal gland (excretory system), hypodermis, and Y-organ (endocrine organ). Our study reports, for the first time in the crustaceans, expression analyses for all four Cas-SODs in hemocytes after immune challenges. Crabs challenged with lipopolysaccharides (LPS) injection had a remarkable induction of Cas-ecCuZnSOD-2 expression along with three other SODs in hemocytes, suggesting that Cas-SODs including Cas-ecCuZnSOD-2 are involved in the defense system, possibly innate immunity and immunocompetency of C. sapidus.
从蓝蟹 Callinectes sapidus 的 EST 文库中最初分离出的全长 cDNA（1362 个核苷酸，GenBank JF736621）编码一种细胞外铜锌超氧化物歧化酶，使用 3' RACE 对其进行了表征，并将其命名为 Cas-ecCuZnSOD-2。 Cas-ecCuZnSOD-2 的开放阅读框包含 203 个推导的氨基酸，具有保守的铜和锌结合的活性催化中心以及两个假定的 N-糖基化和九个磷酸化位点的翻译后修饰。总体而言，Cas-ecCuZnSOD-2 的推导氨基酸与先前在 C. sapidus 中发现的 Cas-ecCuZnSOD（GenBank AF264031）仅共享 35％的序列同一性，而与 Scylla paramamosain ecCuZnSOD（GenBank FJ774661）则显示出约 75％的序列同一性。 Cas-ecCuZnSOD-2 和 C. sapidus 的其他三种 SOD（ecCuZn、cytMn-和 mitMn SOD）的表达谱在从处于蜕皮中期的雌性幼体获得的测试组织中广泛存在：脑，眼柄神经节，围心器和胸神经节复合体（神经系统）；肝胰腺（消化系统）；心脏，动脉和血细胞（循环系统）；鳃和触角腺（排泄系统），皮下组织和 Y 器官（内分泌器官）。我们的研究首次在甲壳动物中报告了 Cas-SODs 在血细胞中的免疫挑战后的表达分析。用脂多糖（LPS）注射对螃蟹进行攻击后，血细胞中 Cas-ecCuZnSOD-2 的表达与其他三种 SOD 一起明显诱导，这表明包括 Cas-ecCuZnSOD-2 在内的 Cas-SODs 参与了防御系统，可能是先天免疫和 C. sapidus 的免疫能力。
