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Cloning and characteristic of MMP1 gene from Hyriopsis cumingii and collagen hydrolytic activity of its recombinant protein

Author:
Hu, Baoqing, Xiao, Jun, Yi, Peipei, Hu, Chenxi, Zhu, Mingxing, Yin, Shuyuan, Wen, Chungen, Wu, Jielian
Source:
Gene 2019 v.693 pp. 92-100
ISSN:
0378-1119
Subject:
Aeromonas hydrophila, Escherichia coli, Hyriopsis cumingii, affinity chromatography, amino acids, angiogenesis, collagen, complementary DNA, gene expression regulation, genes, hemocytes, hepatopancreas, immune response, inflammation, isoelectric point, messenger RNA, metalloproteinases, mice, molecular weight, mussels, peptidoglycans, prototypes, quantitative polymerase chain reaction, rats, recombinant proteins, reverse transcriptase polymerase chain reaction, signal peptide, tissue repair, tissues, transcription (genetics)
Abstract:
Matrix metalloproteinases (MMPs) play an essential role in a variety of biological processes including wound healing, inflammation, cell invasion, angiogenesis and immune defense. In this study, a putative MMP1 cDNA was cloned and characterized from Hyriopsis cumingii (designated as HcMMP1). The cDNA was 1822 bp in length and encoded a putative protein of 510 amino acids, with a predicted molecular mass of 58.28 kDa and an isoelectric point (pI) of 9.27. HcMMP1 contained all prototype MMPs family signatures, such as signal peptide, prodomain, catalytic center, hinge region, and hemopexin like domain. Quantitative real time-PCR (qRT-PCR) revealed that in mussels HcMMP1 mRNA was expressed in all tissues tested, and the transcriptional expression levels were significantly up-regulated in hepatopancreas and hemocytes after Aeromonas hydrophila, peptidoglycan stimulations and in mantle after wounding. Moreover, the recombination HcMMP1 protein, successfully expressed in Escherichia coli, was purified by affinity chromatography with the concentration of final yield at 0.3 mg/mL. The recombinase had an essentially hydrolytic activity toward rat type I collagen, mouse II and IV collagen after renaturation.
Agid:
6303619