Journal
FISH & SHELLFISH IMMUNOLOGY
Volume 119, Issue -, Pages 533-541Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fsi.2021.10.036
Keywords
Tissue inhibitor of metalloproteinase; Matrix metalloproteinases; Wound repair model; Growth rate
Funding
- National Natural Science Foundation of China [13007168, 13007198, 31472305, 21467015, 31460697]
- Support Project of the Modern Agricultural Industry Technology System [ZQT20180027]
- Scientific and Technological in Jiangxi Province [20160BBF60053, 20192BAB204009]
- Nanchang University Seed Grant for Biomedicine of Jiangxi Province, China
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In this study, TIMP-1, 2 from Hyriopsis cumingii were cloned and identified, with potential involvement in the immune response and efficient expression in Escherichia coli. The findings suggest that HcTIMP-1, 2 can inhibit metalloproteinase activity and promote cell growth.
Metalloproteinase tissue inhibitors (TIMPs) have the activity of inhibiting matrix metalloproteinases (MMPs), which can promote cell growth, bind to the matrix, inhibit angiogenesis, and play a key role in extracellular matrix (ECM) metabolism regulation. In this study, TIMP-1, 2 from Hyriopsis cumingii (designated as HcTIMP-1, 2) were cloned and identified. Full-length cDNA of HcTIMP-1, 2 was 1160 bp and 729 bp, encoding 235 and 150 amino acid residues, respectively. The predicted molecular weight of HcTIMP-1 and 2 protein was 27.26 and 16.58 kDa, with isoelectric points of 8.89 and 8.72, respectively. HcTIMP-2 contained only one netrin (NTR) domain at the N-terminal but lacked a C-terminal domain. The mRNA of HcTIMP-1, 2 was expressed in hepatopancreas, gills, muscles, hemocytes, and mantles, which had the highest expression in hemocytes and muscles. The expression of HcTIMP-1, 2 had increased remarkably in hemocytes after bacterial challenge. After trauma, HcTIMP-1, 2 genes had the highest expression level in the first day. This indicated that HcTIMP-1 and 2 were involved in the immune response of H. cumingii. The soluble recombinant proteins HcTIMP-1, 2 were expressed efficiently in Escherichia coli BL21 (DE3) by constructing pET32a-TIMP1, 2 recombinant plasmids. The concentration of the recombinant was 0.14 and 0.31 mg/mL, respectively. The recombinant HcTIMP-1, 2 proteins were shown to inhibit human MMP2 activity and promoted the growth of NBL-7 and HUVE cells.
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