Song Weiwei, Mu Changkao, Li Ronghua, Wang Chunlin
Key Laboratory of Applied Marine Biotechnology (Ningbo University), Ministry of Education, Ningbo, 315211, China; Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo 315211, China.
Key Laboratory of Applied Marine Biotechnology (Ningbo University), Ministry of Education, Ningbo, 315211, China; Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo 315211, China.
Fish Shellfish Immunol. 2017 Aug;67:596-603. doi: 10.1016/j.fsi.2017.06.034. Epub 2017 Jun 12.
The aim of this work was constructive to understand the function of peroxiredoxin (PRDX) family member Peroxiredoxin 1 in Sepiella maindroni (SmPrx1) through molecular mechanisms of reproduction, embryonic development and immune responses to Vibrio alginolyticus. The full-length cDNA of SmPrx1 was of 1062 bp, contains a 5' untranslated region (UTR) of 79bp, a 3' UTR of 359 bp, an open reading frame of 624 bp encoding 207 amino acids. The conserved peroxidase catalytic center "FYPLDFTFVCPTEI" and "GEVCPA" were observed in the sequence of SmPrx1; this indicated that it was a member of 2-Cys Prx. Quantitative real-time (qRT)-PCR assays revealed that SmPrx1 was ubiquitously expressed in all examined tissues, muscle, ink sac, liver, ovary, testis, intestine, gill and totally blood cells, and showed high levels in testis. SmPrx1 mRNA was ubiquitously detected in all tested tissues, and the expression was comparatively high in testis, hemocyte, liver and ovary. Moreover, the SmPrx1 gene transcript was detected at all five stages of embryonic development phases that were respectively the zygote stage, the pre-embryonic stage, the organogenesis stage, the morphological integrity stage, the pre-hatching stage. The general tendency of expression was gradually increased and rapidly decreased. High expressed in progenitive tissues and embryonic development exhibit the proliferation-associated protein characterization like in mammal. The expression levels of SmPrx1 in liver and hemocytes grew swiftly and quickly reached peak value after Vibrio alginolyticus challenge. As hours passed by, the expression level began to reduce and resumed to normal levels after 48 h. The antioxidant activity and peroxidase activity of SmPrx1 were 6.17 U/mg. The results showed that the recombined protein of SmPrx1 had antioxidant activity and was the importance part of the antioxidant system in Sepiella maindroni. This study provides useful information to help further understand the functional mechanism of Prx 1 in marine cephalopod immunity.
这项工作的目的是通过繁殖、胚胎发育以及对溶藻弧菌免疫反应的分子机制，来深入了解过氧化物酶（PRDX）家族成员——曼氏无针乌贼过氧化物酶1（SmPrx1）的功能。SmPrx1的全长cDNA为1062 bp，包含一个79 bp的5'非翻译区（UTR）、一个359 bp的3'UTR以及一个624 bp的开放阅读框，编码207个氨基酸。在SmPrx1序列中观察到保守的过氧化物酶催化中心“FYPLDFTFVCPTEI”和“GEVCPA”；这表明它是2-Cys Prx的成员。实时定量（qRT）-PCR分析显示，SmPrx1在所有检测组织（肌肉、墨囊、肝脏、卵巢、睾丸、肠道、鳃和全血细胞）中均有广泛表达，且在睾丸中表达水平较高。SmPrx1 mRNA在所有测试组织中均能检测到，在睾丸、血细胞、肝脏和卵巢中的表达相对较高。此外，在胚胎发育的所有五个阶段（分别为合子期、胚胎前期、器官发生期、形态完整性期、孵化前期）均检测到SmPrx1基因转录本。表达的总体趋势是逐渐升高然后迅速下降。在生殖组织和胚胎发育中高表达，表现出与哺乳动物中增殖相关蛋白类似的特征。溶藻弧菌攻击后，肝脏和血细胞中SmPrx1的表达水平迅速升高，并在短时间内达到峰值。随着时间的推移，表达水平开始下降，并在48小时后恢复到正常水平。SmPrx1的抗氧化活性和过氧化物酶活性为6.17 U/mg。结果表明，SmPrx1重组蛋白具有抗氧化活性，是曼氏无针乌贼抗氧化系统的重要组成部分。本研究为进一步了解Prx 1在海洋头足类动物免疫中的功能机制提供了有用信息。
