Wang Xian-Wei, Xu Wen-Teng, Zhang Xiao-Wen, Zhao Xiao-Fan, Yu Xiao-Qiang, Wang Jin-Xing
The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education, School of Life Sciences, Shandong University, Jinan, Shandong 250100, PR China.
Fish Shellfish Immunol. 2009 Oct;27(4):556-62. doi: 10.1016/j.fsi.2009.07.011. Epub 2009 Jul 31.
C-type lectins may function as pattern-recognition receptors (PRRs) and play important roles in immune responses. In this work, a cDNA for a new C-type lectin, FcLec3, was obtained from Chinese white shrimp Fenneropenaeus chinensis using expressed sequence tag analysis and rapid amplification of the cDNA ends. FcLec3 contains an N-terminal signal peptide and a carbohydrate recognition domain (CRD). RT-PCR analysis showed that FcLec3 was mainly expressed in hepatopancreas and that the expression of FcLec3 was obviously up-regulated by Vibrio anguillarum or white spot syndrome virus (WSSV) challenge. Recombinant FcLec3 could agglutinate Gram-negative and -positive bacteria with the presence of calcium. A following agglutination inhibitory test indicated that FcLec3 could recognize muramic acid and peptidoglycan. Besides, pull-down assay showed that the recombinant protein could interact with VP28, one major envelope protein of WSSV. These results suggested that FcLec3 might function in the recognition of bacterial and viral pathogens in shrimp.
C型凝集素可能作为模式识别受体(PRRs),并在免疫反应中发挥重要作用。在本研究中,利用表达序列标签分析和cDNA末端快速扩增技术,从中国对虾凡纳滨对虾中获得了一种新的C型凝集素FcLec3的cDNA。FcLec3包含一个N端信号肽和一个碳水化合物识别结构域(CRD)。RT-PCR分析表明,FcLec3主要在肝胰腺中表达,鳗弧菌或白斑综合征病毒(WSSV)刺激后,FcLec3的表达明显上调。重组FcLec3在有钙存在的情况下能够凝集革兰氏阴性菌和阳性菌。随后的凝集抑制试验表明,FcLec3能够识别胞壁酸和肽聚糖。此外,下拉试验表明,重组蛋白能够与WSSV的一种主要包膜蛋白VP28相互作用。这些结果表明,FcLec3可能在对虾识别细菌和病毒病原体中发挥作用。