Target-directed isolation and identification of a serum lectin from lamprey (Lampetra japonica) by chromatographys and MALDI-TOF/TOF  

作　　者：HAN Yinglun GOU Meng SONG Xiaoping SONG Tao SHI Biyue PANG Yue LI Qingwei 

机构地区：[1]College of Life Science, Liaoning Normal University, Dalian 116029, China [2]Lamprey Research Center, Liaoning Normal University, Dalian 116029, China [3]Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China

出　　处：《Acta Oceanologica Sinica》2018年第5期113-116,共4页海洋学报（英文版）

基　　金：The National Program on Key Basic Research Project(973 Program)of China under contract No.2013CB835304;the National Marine Public Projects under contract No.201305016;the National Natural Science Foundation of China under contract Nos31772884 and 31601865;the Key Projects of Scientific Research Platform of Liaoning Provincial Education Department under contract No.L201683651

摘　　要：A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.

关 键 词：Lampetra japonica lectin purification identification MALDI-TOF/TOF 

分 类 号：Q51[生物学—生物化学]