An interaction between leukocyte cell-derived chemotaxin 2 and transferrin of ayu, Plecoglossus altivelis
An interaction between leukocyte cell-derived chemotaxin 2 and transferrin of ayu, Plecoglossus altivelis
复制标题
DOI：
10.1016/j.fsi.2009.02.010
复制
发表时间：
2009-03-01
影响因子：
4.7
通讯作者：
Li, Ming Y.
中科院分区：
文献类型：
--
作者：
Chen, Jiong;Yang, Hong Y.;Li, Ming Y.
利用酵母双杂交（Y2 H）技术筛选香鱼肝脏cDNA表达文库与白细胞趋化因子2（LECT-2）的相互作用。在鉴定的9个独立相互作用克隆中，7个是相同的，并且与鱼类的转铁蛋白（To）基因密切相关，而另外两个与c型凝集素基因相关。香鱼Tf（aTf）和香鱼LECT 2（aLECT 2）之间的相互作用通过两种蛋白的体外共免疫沉淀来证实。使用两种蛋白质不同部分的Y2 H测定表明，aTf 185 -289片段不参与与成熟aLECT 2的相互作用，而aLECT 2的转运肽不能与整个aTf相互作用。计算机分析表明，aTf 185 -289含有两个铁结合残基Tyr 197和His 253，位于aTf N端。LECT 2与来自同一动物的Tf之间也确定了强相互作用，例如大黄鱼、大黄鱼和小鼠、小家鼠。然而，未确定跨种属相互作用。基于已发表的数据，Tf-LECT 2相互作用被认为最有可能参与机体对感染的防御。(C)2009爱思唯尔有限公司保留所有权利。
Yeast two-hybrid (Y2H) screens were used to test for interactions between leukocyte cell-derived chemotaxin 2 (LECT-2) and a liver cDNA expression library of ayu, Plecoglossus altivelis. Of the 9 independent interacting clones identified, 7 were identical and closely related to transferrin (To genes of fish, while the other two were related to c-type lectin genes. The interaction between ayu Tf (aTf) and ayu LECT2 (aLECT2) was confirmed by in vitro co-immunoprecipitation of the two proteins. Y2H assays using different parts of the two proteins showed that the segment aTf185-289 was not involved in the interaction with mature aLECT2, while the transit peptide of aLECT2 couldn't interact with entire aTf. Computer analysis revealed that aTf185-289, which contained two iron binding residues, Tyr197 and His253, was located at the N-terminus of aTf N-lobe. Strong interactions were also determined between LECT2 and Tf from the same animal, such as croceine croaker, Larimichthys crocea and mouse, Mus musculus. However, no cross-species interactions were determined. Based on published data, the Tf-LECT2 interaction is suggested to be most possibly involved in the body's defense against infection. (C) 2009 Elsevier Ltd. All rights reserved.