Involvement of a Serpin serine protease inhibitor (OoSerpin) from mollusc Octopus ocellatus in antibacterial response.
Involvement of a Serpin serine protease inhibitor (OoSerpin) from mollusc Octopus ocellatus in antibacterial response.
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DOI：
10.1016/j.fsi.2014.10.028
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发表时间：
2015
影响因子：
4.7
通讯作者：
Xiumei Wei;Jie Xu;Jianmin Yang;Xiangquan Liu;Ranran Zhang;Weijun Wang;Jialong Yang
中科院分区：
文献类型：
--
作者：
Xiumei Wei;Jie Xu;Jianmin Yang;Xiangquan Liu;Ranran Zhang;Weijun Wang;Jialong Yang
丝氨酸蛋白酶抑制剂（SPIs）是丝氨酸蛋白酶抑制剂（serine protease inhibitors, SPIs）的重要成员，能够调控蛋白水解事件，参与多种生理过程。本研究从ococtopus ocellatus（命名为OoSerpin）中鉴定出一个Serpin同源物。OoSerpin全长1735 bp，包含214 bp的5 ‘未翻译区，282 bp的3 ’ UTR和1239 bp的开放阅读框。该开放阅读框编码了一个412个氨基酸的多肽，预测分子量为46.5 kDa，等电点为8.52。OoSerpin蛋白与来自musus musus （NP_941373）和dixodes肩胛骨（XP_002407493）的其他serpin蛋白序列同源性为37%。保守SERPIN结构域的存在强烈表明OoSerpin是SERPIN亚家族的成员。然后对OoSerpin在组织和细菌刺激下的表达模式进行了表征。OoSerpin mRNA在未处理edo的所有组织中均有不同水平的组成性表达。眼状体，包括地幔（最低）、肌肉、肾囊、鳃、血细胞、性腺、全身心脏和肝胰腺（最高）。在对照组中，OoSerpin的转录水平显著上调（P< 0.01）。鳗弧菌和黄体微球菌对细菌的攻击，表明其参与了抗菌免疫反应。此外，OoSerpin的重组蛋白rOoSerpin表现出较强的抑制胰蛋白酶和凝乳胰蛋白酶活性以及大肠杆菌生长的能力。我们的研究结果表明，OoSerpin是一种潜在的抗菌因子，参与了ofO的免疫反应。抗细菌感染。
Serpin is an important member of serine protease inhibitors (SPIs), which is capable of regulating proteolytic events and involving in a variety of physiological processes. In present study, a Serpin homolog was identified fromOctopus ocellatus(designated as OoSerpin). Full-length cDNA of OoSerpin was of 1735 bp, containing a 5′ untranslated region of 214 bp, a 3′ UTR of 282 bp, and an open reading frame of 1239 bp. The open reading frame encoded a polypeptide of 412 amino acids which has a predicted molecular weight of 46.5 kDa and an isoelectric point of 8.52. The OoSerpin protein shares 37% sequence identity with other Serpins fromMus musculus(NP_941373) andIxodes scapularis(XP_002407493). The existence of a conserved SERPIN domain strongly suggested that OoSerpin was a member of the Serpin subfamily. Expression patterns of OoSerpin, both in tissues and towards bacterial stimulation, were then characterized. The mRNA of OoSerpin was constitutively expressed at different levels in all tested tissues of untreatedO.ocellatus, including mantle (lowest), muscle, renal sac, gill, hemocyte, gonad, systemic heart, and hepatopancreas (highest). The transcriptional level of OoSerpin was significantly up-regulated (P< 0.01) inO.ocellatusupon bacterial challenges withVibrio anguillarumandMicrococcus luteus, indicating its involvement in the antibacterial immune response. Furthermore, rOoSerpin, the recombinant protein of OoSerpin, exhibited strong abilities to inhibit proteinase activities of trypsin and chymotrypsin as well as the growth ofEscherichia coli. Our results demonstrate that OoSerpin is a potential antibacterial factor involved in the immune response ofO.ocellatusagainst bacterial infection.