Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas

doi:10.1016/j.dci.2018.08.014

IOCAS-IR

	Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas
	Lv, Zhao 1,2,3; Qiu, Limei 1; Liu, Zhaoqun 2,4; Wang, Weilin 2,4; Chen, Hao 1; Jia, Yunke 1,2,3; Jia, Zhihao 1,2,3; Jiang, Shuai 1; Wang, Lingling 2,4; Song, Linsheng 2,4
	2018-12-01
发表期刊	DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN	0145-305X
卷号	89 页码:152-162
通讯作者	Song, Linsheng(lshsong@dlou.edu.cn)
摘要	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.
关键词	Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker
DOI	10.1016/j.dci.2018.08.014
收录类别	SCI
语种	英语
资助项目	Talented Scholars in Dalian Ocean University ; Research Foundation for Distinguished professor of Liaoning ; Earmarked Fund from Modern Agro-industry Technology Research System[CARS-49] ; Qingdao National Laboratory for Marine Science and Technology[2017ASTCP-OS13] ; AoShan Talents Cultivation Program ; Dalian High Level Talent Innovation Support Program[2015R020] ; National Natural Science Foundation of China[U1706204] ; 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研究方向	Fisheries ; Immunology ; Zoology
WOS类目	Fisheries ; Immunology ; Zoology
WOS记录号	WOS:000444932200017
出版者	ELSEVIER SCI LTD
WOS关键词	L-LIKE PROTEINASES ; CYSTEINE CATHEPSINS ; ANTIGEN PRESENTATION ; GENE-EXPRESSION ; LARVAL MIDGUT ; HEMOCYTES ; IMMUNITY ; ACTIVATION ; PEPTIDASES ; BACTERIAL
引用统计
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/160234
通讯作者	Song, Linsheng
作者单位	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
推荐引用方式 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.