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| Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas | |
Lv, Zhao1,2,3; Qiu, Limei1; Liu, Zhaoqun2,4 ; Wang, Weilin2,4; Chen, Hao1; Jia, Yunke1,2,3; Jia, Zhihao1,2,3; Jiang, Shuai1; Wang, Lingling2,4; Song, Linsheng2,4
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| 2018-12-01 | |
| Source Publication | DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
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| ISSN | 0145-305X |
| Volume | 89Pages:152-162 |
| Corresponding Author | Song, Linsheng(lshsong@dlou.edu.cn) |
| Abstract | Cathepsin L1 (CTSL1) is a lysosomal cysteine protease with a papain-like structure. It is known to be implicated in multiple processes of immune response against pathogen infection based on the proteolytic activity. In the present study, a CTSL1 homologue (designated as CgCTSL1) was identified from Crassostrea gigas. It contained a typically single Pept_C1 domain with three conserved catalytically essential residues (Gln(25), His(135) and Asn(125) ). The mRNA of CgCTSL1 was ubiquitously expressed in oyster tissues with the highest expression level in important immune tissues such as gill and hemocytes. CgCTSL1 proteins were mainly detected in gill and hepatopancreas by immunohistochemistry. Recombinant CgCTSL1 (rCgCTSL1) exhibited proteolytic activity to cleave the substrate Ac-FR-amino-4-trifiuoromethyl coumarin (AFC) in a dose-dependent manner, and the inhibitor could reduce its proteolytic activity. After the interference of CgCTSL1 mRNA, the proteolytic activity of oyster hemocytes was significantly down-regulated with the released AFC fluorescence value decreasing from 375.84 to 179.21 (p < 0.05). Flow cytometry analysis revealed that the expression of CgCTSL1 protein was higher in phagocytes with the mean fluorescence intensity (MFI) value of 21,187 (4.13-fold, p < 0.01) compared to the MFI value of 5,130 in non-phagocytic hemocytes. The further confocal analysis demonstrated that the actively phagocytic hemocytes with green bead signals were co-localized with stronger CgCTSL1 positive signals. The mRNA expression levels of CgCTSL1 in phagocyte-like sub-populations of granulocytes and semi-granulocytes were 298.12-fold (p < 0.01) and 2.75-fold (p < 0.01) of that in agranulocytes, respectively. Western blotting analysis of the hemocyte proteins revealed that CgCTSL1 was relatively abundant in granulocytes and semigranulocytes compared to that in agranulocytes. These results collectively suggested that CgCTSL1, a CTSL1 homologue highly expressed in phagocyte-like hemocytes, was possibly involved in cellular immune response dependent on its conserved proteolytic activity, which might provide clues for the divergence between phagocytes and non-phagocytic hemocytes as well as the identification of promising molecular markers for phagocytes in oyster C. gigas. |
| Keyword | Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker |
| DOI | 10.1016/j.dci.2018.08.014 |
| Indexed By | SCI |
| Language | 英语 |
| Funding Project | National Natural Science Foundation of China[U1706204] ; Dalian High Level Talent Innovation Support Program[2015R020] ; AoShan Talents Cultivation Program ; Qingdao National Laboratory for Marine Science and Technology[2017ASTCP-OS13] ; Earmarked Fund from Modern Agro-industry Technology Research System[CARS-49] ; Research Foundation for Distinguished professor of Liaoning ; Talented Scholars in Dalian Ocean University ; National Natural Science Foundation of China[U1706204] ; Dalian High Level Talent Innovation Support Program[2015R020] ; AoShan Talents Cultivation Program ; Qingdao National Laboratory for Marine Science and Technology[2017ASTCP-OS13] ; earmarked fund from Modern Agro-industry Technology Research System[CARS-49] ; Research Foundation for Distinguished professor of Liaoning ; Talented Scholars in Dalian Ocean University |
| WOS Research Area | Fisheries ; Immunology ; Zoology |
| WOS Subject | Fisheries ; Immunology ; Zoology |
| WOS ID | WOS:000444932200017 |
| Publisher | ELSEVIER SCI LTD |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.qdio.ac.cn/handle/337002/160236 |
| Collection | 实验海洋生物学重点实验室 |
| Corresponding Author | Song, Linsheng |
| Affiliation | 1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266235, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Dalian Ocean Univ, Liaoning Key Lab Marine Anim Immunol & Dis Contro, Dalian 116023, Peoples R China |
| First Author Affilication | Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences |
| Recommended Citation GB/T 7714 | Lv, Zhao,Qiu, Limei,Liu, Zhaoqun,et al. Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2018,89:152-162. |
| APA | Lv, Zhao.,Qiu, Limei.,Liu, Zhaoqun.,Wang, Weilin.,Chen, Hao.,...&Song, Linsheng.(2018).Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,89,152-162. |
| MLA | Lv, Zhao,et al."Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 89(2018):152-162. |
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| Molecular characteri（3509KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | View | |
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