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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
2018-12-01
Source PublicationDEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN0145-305X
Volume89Pages:152-162
Corresponding AuthorSong, Linsheng(lshsong@dlou.edu.cn)
AbstractCathepsin 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.
KeywordCrassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker
DOI10.1016/j.dci.2018.08.014
Indexed BySCI
Language英语
Funding ProjectNational 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 AreaFisheries ; Immunology ; Zoology
WOS SubjectFisheries ; Immunology ; Zoology
WOS IDWOS:000444932200017
PublisherELSEVIER SCI LTD
Citation statistics
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/160236
Collection实验海洋生物学重点实验室
Corresponding AuthorSong, Linsheng
Affiliation1.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 AffilicationKey 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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