A lectin (CfLec-2) aggregating Staphylococcus haemolyticus from scallop Chlamys farreri.
A lectin (CfLec-2) aggregating Staphylococcus haemolyticus from scallop Chlamys farreri.
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DOI：
10.1016/j.fsi.2007.11.014
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发表时间：
2008-03
影响因子：
4.7
通讯作者：
P. Zheng;Hao Wang;Jianmin Zhao;L. Song;L. Qiu;Chaohua Dong;Bo Wang;Yunchao Gai;C. Mu;Chenghua Li;Duojiao Ni;K. Xing
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文献类型：
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作者：
P. Zheng;Hao Wang;Jianmin Zhao;L. Song;L. Qiu;Chaohua Dong;Bo Wang;Yunchao Gai;C. Mu;Chenghua Li;Duojiao Ni;K. Xing
凝集素是一个碳水化合物识别蛋白家族，在先天免疫中起着至关重要的作用。本研究通过EST和RACE方法从栉孔扇贝中克隆了一个新的凝集素基因CfLec-2。CfLec-2的全长cDNA由708bp组成，编码130个残基的典型的长碳水化合物识别结构域。推导的氨基酸序列与智人短链蛋白、鳗鱼C型凝集素-1和凝集素-2有很高的相似性。将编码CfLec-2成熟肽的基因片段重组到表达载体pET-32a(+)中，并在大肠杆菌Rosseta-Gami(DE3)中表达。重组CfLec-2(rCfLec-2)蛋白对溶血葡萄球菌具有凝集活性，其凝集活性可被d-甘露糖抑制，但不能被EDTA或d-半乳糖抑制，表明CfLec-2是一种不依赖于钙离子的凝集素。此外，rCfLec-2还能抑制大肠杆菌TOP10F‘的生长。这些结果表明，CfLec-2可能参与了对扇贝体内细菌病原体的识别和清除。
Lectins are a family of carbohydrate-recognition proteins which play crucial roles in innate immunity. In this study, a new lectin (CfLec-2) gene was cloned from Chlamys farreri by EST and RACE approaches. The full-length cDNA of CfLec-2 was composed of 708bp, encoding a typical long form carbohydrate-recognition domain of 130 residues. The deduced amino acid sequence showed high similarity to Brevican in Homo sapiens, C-type lectin-1 and lectin-2 in Anguilla japonica. The cDNA fragment encoding the mature peptide of CfLec-2 was recombined into plasmid pET-32a (+) and expressed in Escherichia coli Rosseta-Gami (DE3). The recombinant CfLec-2 (rCfLec-2) protein exhibited aggregative activity toward Staphylococcus haemolyticus, and the agglutination could be inhibited by d-mannose but not EDTA or d-galactose, indicating that CfLec-2 was a Ca2+independent lectin. Moreover, rCfLec-2 could suppress the growth of E. coli TOP10F′. These results suggested that CfLec-2 was perhaps involved in the recognition and clearance of bacterial pathogens in scallop.