Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas
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...
Published in: | Developmental & Comparative Immunology |
---|---|
Main Authors: | , , , , , , , , , |
Format: | Report |
Language: | English |
Published: |
ELSEVIER SCI LTD
2018
|
Subjects: | |
Online Access: | http://ir.qdio.ac.cn/handle/337002/160234 https://doi.org/10.1016/j.dci.2018.08.014 |
id |
ftchinacasciocas:oai:ir.qdio.ac.cn:337002/160234 |
---|---|
record_format |
openpolar |
spelling |
ftchinacasciocas:oai:ir.qdio.ac.cn:337002/160234 2023-05-15T15:57:57+02:00 Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas Lv, Zhao Qiu, Limei Liu, Zhaoqun Wang, Weilin Chen, Hao Jia, Yunke Jia, Zhihao Jiang, Shuai Wang, Lingling Song, Linsheng 2018-12-01 http://ir.qdio.ac.cn/handle/337002/160234 https://doi.org/10.1016/j.dci.2018.08.014 英语 eng ELSEVIER SCI LTD DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY http://ir.qdio.ac.cn/handle/337002/160234 doi:10.1016/j.dci.2018.08.014 Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker Fisheries Immunology Zoology L-LIKE PROTEINASES CYSTEINE CATHEPSINS ANTIGEN PRESENTATION GENE-EXPRESSION LARVAL MIDGUT HEMOCYTES IMMUNITY ACTIVATION PEPTIDASES BACTERIAL 期刊论文 2018 ftchinacasciocas https://doi.org/10.1016/j.dci.2018.08.014 2022-06-27T05:40:20Z 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 ... Report Crassostrea gigas Pacific oyster Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR Pacific Developmental & Comparative Immunology 89 152 162 |
institution |
Open Polar |
collection |
Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR |
op_collection_id |
ftchinacasciocas |
language |
English |
topic |
Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker Fisheries Immunology Zoology L-LIKE PROTEINASES CYSTEINE CATHEPSINS ANTIGEN PRESENTATION GENE-EXPRESSION LARVAL MIDGUT HEMOCYTES IMMUNITY ACTIVATION PEPTIDASES BACTERIAL |
spellingShingle |
Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker Fisheries Immunology Zoology L-LIKE PROTEINASES CYSTEINE CATHEPSINS ANTIGEN PRESENTATION GENE-EXPRESSION LARVAL MIDGUT HEMOCYTES IMMUNITY ACTIVATION PEPTIDASES BACTERIAL Lv, Zhao Qiu, Limei Liu, Zhaoqun Wang, Weilin Chen, Hao Jia, Yunke Jia, Zhihao Jiang, Shuai Wang, Lingling Song, Linsheng Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
topic_facet |
Crassostrea gigas Phagocyte-highly expressed Cathepsin L1 Proteolytic activity Molecular marker Fisheries Immunology Zoology L-LIKE PROTEINASES CYSTEINE CATHEPSINS ANTIGEN PRESENTATION GENE-EXPRESSION LARVAL MIDGUT HEMOCYTES IMMUNITY ACTIVATION PEPTIDASES BACTERIAL |
description |
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 ... |
format |
Report |
author |
Lv, Zhao Qiu, Limei Liu, Zhaoqun Wang, Weilin Chen, Hao Jia, Yunke Jia, Zhihao Jiang, Shuai Wang, Lingling Song, Linsheng |
author_facet |
Lv, Zhao Qiu, Limei Liu, Zhaoqun Wang, Weilin Chen, Hao Jia, Yunke Jia, Zhihao Jiang, Shuai Wang, Lingling Song, Linsheng |
author_sort |
Lv, Zhao |
title |
Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
title_short |
Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
title_full |
Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
title_fullStr |
Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
title_full_unstemmed |
Molecular characterization of a cathepsin L1 highly expressed in phagocytes of pacific oyster Crassostrea gigas |
title_sort |
molecular characterization of a cathepsin l1 highly expressed in phagocytes of pacific oyster crassostrea gigas |
publisher |
ELSEVIER SCI LTD |
publishDate |
2018 |
url |
http://ir.qdio.ac.cn/handle/337002/160234 https://doi.org/10.1016/j.dci.2018.08.014 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_relation |
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY http://ir.qdio.ac.cn/handle/337002/160234 doi:10.1016/j.dci.2018.08.014 |
op_doi |
https://doi.org/10.1016/j.dci.2018.08.014 |
container_title |
Developmental & Comparative Immunology |
container_volume |
89 |
container_start_page |
152 |
op_container_end_page |
162 |
_version_ |
1766393655266902016 |