
在仿刺参体腔液中发现新的调理素样分子
A new opsonin-like protein identified from coelomic fluid of sea cucumber Apostichopus japonicus
Mai Kangsen、Meng Fanyi、Ma Hongming、Zhang Wenbing
通过研究发现,仿刺参体腔液中存在一种新的调理素样分子。通过SDS-PAGE,凝胶过滤层析,质谱分析等方法进行蛋白质的纯化和分析,采用荧光显微镜观察吞噬细胞对酵母细胞的吞噬作用,发现一种分子量约为18 kDa的分子可以结合到酵母细胞表面,并可以增强吞噬细胞的吞噬作用。推测这种分子是无脊椎动物调理素样分子中的新成员。同时也发现,虽然调理素样分子能够增强吞噬细胞的吞噬作用,但是,它不是单独起作用的,仍然必须依赖体腔液中其他未知成分,而对未知成分的鉴定需要进一步研究。
In order to determine if opsonin-like molecules exist in the coelomic fluid of sea cucumber Apostichopus japonicus and identify their properties, immunochemical methods including phagocytosis assay, SDS-PAGE, gel filtration and mass spectrometric analysis were performed. The phagocytosis of sea cucumber phagocytes was quantified in vitro using heat-killed yeast cells as target cells. Phagocytes of the sea cucumber were capable of phagocytosis in isotonic buffer, cell-free coelomic fluid (CFCF) and yeast cell-adsorbed CFCF, but the phagocytosis was significantly enhanced when the incubation medium was CFCF. This phagocytosis promoting activity of CFCF was eliminated by pre-adsorption of CFCF with yeast cells. SDS-PAGE analysis showed that a protein with 18 kDa was able to bind to yeast cells. By using gel filtration, the protein was isolated from coelomic fluid of the sea cucumber. Functional test of this molecule demonstrated that it was a phagocytosis-enhancing opsonin-like molecule. However, the phagocytosis-enhancing function of the opsonin-like molecule was dependent on some unknown factor(s) in the CFCF, which needs further investigations. Mass spectrometric analysis indicated that this opsonin-like molecule is a new protein.
postichopus japonicus / phagocyte / coelomic fluid / opsonin-like molecule
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