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| Identification and characterization of a serine protease inhibitor Esserpin from the Chinese mitten crab Eriocheir sinensis | |
| Wang, Lingling1; Ma, Zhaopeng1,2; Yang, Jialong1; Gai, Yunchao1; Zhou, Zhi1; Wang, Leilei1,2; Yue, Feng1,2; Song, Linsheng1; Wang, LL | |
| 2013-06-01 | |
| 发表期刊 | FISH & SHELLFISH IMMUNOLOGY
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| ISSN | 1050-4648 |
| 卷号 | 34期号:6页码:1576-1586 |
| 文章类型 | Article |
| 摘要 | Serine protease inhibitors (serpins) represent an expanding superfamily of endogenous inhibitors that regulate proteolytic events and involve in a variety of physiological processes. A serine protease inhibitor, namely Esserpin, was identified from Chinese mitten crab Eriocheir sinensis based on expressed sequence tag (EST) analysis. The full-length cDNA of Esserpin was of 2367 bp, including an open reading frame (ORE) of 1371 bp encoding a polypeptide of 456 amino acids with estimated molecular mass of 49.95 kDa and theoretical isoelectric point of 6.03. A putative signal peptide of 23 amino acids and a classical serpin domain were identified in Esserpin. The deduced amino acid sequence of Esserpin shared homology with serpins from Fenneropenaeus chinensis and Pacifastacus leniusculus. The mRNA transcripts of Esserpin could be detected in all the examined tissues including heart, gill, hemocytes, muscle, gonad and hepatopancreas, and the highest expression level was present in gonad. After the crabs were challenged by Vibrio anguillarum and Pichia pastoris, the expression levels of Esserpin transcripts in hemocytes were significantly up-regulated, and peaked at 24 h (5.18-fold of blank group, P < 0.05) and 3 h (2.87-fold of blank group, P < 0.05), respectively. The functional activity of Esserpin was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli BL21 (DE3)-pLysS. The recombinant Esserpin (rEsserpin) could inhibit trypsin activities in a dose-dependent manner, and it could lead to 100% inhibition of trypsin activities under the concentration of 873.76 nM, while there was no evident inhibition of chymotrypsin observed with rEsserpin. Moreover, rEsserpin inhibited the growth of E. coli at the final concentration of 1747.52 nM, and it also significantly depressed (P < 0.05) the phenoloxidase activity in the plasma at the final concentration of 873.76 nM. These results indicated that Esserpin was a homologue of serpin in crab and it could be induced after immune stimulation and mediate immune response possibly via the inhibition of bacterial growth and the regulation of prophenoloxidase-activating system. (C) 2013 Elsevier Ltd. All rights reserved.; Serine protease inhibitors (serpins) represent an expanding superfamily of endogenous inhibitors that regulate proteolytic events and involve in a variety of physiological processes. A serine protease inhibitor, namely Esserpin, was identified from Chinese mitten crab Eriocheir sinensis based on expressed sequence tag (EST) analysis. The full-length cDNA of Esserpin was of 2367 bp, including an open reading frame (ORE) of 1371 bp encoding a polypeptide of 456 amino acids with estimated molecular mass of 49.95 kDa and theoretical isoelectric point of 6.03. A putative signal peptide of 23 amino acids and a classical serpin domain were identified in Esserpin. The deduced amino acid sequence of Esserpin shared homology with serpins from Fenneropenaeus chinensis and Pacifastacus leniusculus. The mRNA transcripts of Esserpin could be detected in all the examined tissues including heart, gill, hemocytes, muscle, gonad and hepatopancreas, and the highest expression level was present in gonad. After the crabs were challenged by Vibrio anguillarum and Pichia pastoris, the expression levels of Esserpin transcripts in hemocytes were significantly up-regulated, and peaked at 24 h (5.18-fold of blank group, P < 0.05) and 3 h (2.87-fold of blank group, P < 0.05), respectively. The functional activity of Esserpin was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli BL21 (DE3)-pLysS. The recombinant Esserpin (rEsserpin) could inhibit trypsin activities in a dose-dependent manner, and it could lead to 100% inhibition of trypsin activities under the concentration of 873.76 nM, while there was no evident inhibition of chymotrypsin observed with rEsserpin. Moreover, rEsserpin inhibited the growth of E. coli at the final concentration of 1747.52 nM, and it also significantly depressed (P < 0.05) the phenoloxidase activity in the plasma at the final concentration of 873.76 nM. These results indicated that Esserpin was a homologue of serpin in crab and it could be induced after immune stimulation and mediate immune response possibly via the inhibition of bacterial growth and the regulation of prophenoloxidase-activating system. (C) 2013 Elsevier Ltd. All rights reserved. |
| 关键词 | Eriocheir Sinensis Serpin Microbial Challenge Antibacterial Activity Prophenoloxidase-activating System |
| 学科领域 | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
| DOI | 10.1016/j.fsi.2013.03.371 |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
| WOS类目 | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
| WOS记录号 | WOS:000319646100022 |
| WOS关键词 | SHRIMP PENAEUS-MONODON ; REACTIVE CENTER LOOP ; MANDUCA-SEXTA ; PROTEINASE-INHIBITOR ; PROPHENOLOXIDASE ACTIVATION ; RECOGNITION PROTEIN ; SERPIN SUPERFAMILY ; GENE-EXPRESSION ; CDNA CLONING ; BLOOD-CELLS |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/16588 |
| 专题 | 实验海洋生物学重点实验室 |
| 通讯作者 | Wang, LL |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
| 第一作者单位 | 实验海洋生物学重点实验室 |
| 推荐引用方式 GB/T 7714 | Wang, Lingling,Ma, Zhaopeng,Yang, Jialong,et al. Identification and characterization of a serine protease inhibitor Esserpin from the Chinese mitten crab Eriocheir sinensis[J]. FISH & SHELLFISH IMMUNOLOGY,2013,34(6):1576-1586. |
| APA | Wang, Lingling.,Ma, Zhaopeng.,Yang, Jialong.,Gai, Yunchao.,Zhou, Zhi.,...&Wang, LL.(2013).Identification and characterization of a serine protease inhibitor Esserpin from the Chinese mitten crab Eriocheir sinensis.FISH & SHELLFISH IMMUNOLOGY,34(6),1576-1586. |
| MLA | Wang, Lingling,et al."Identification and characterization of a serine protease inhibitor Esserpin from the Chinese mitten crab Eriocheir sinensis".FISH & SHELLFISH IMMUNOLOGY 34.6(2013):1576-1586. |
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