Target-directed isolation and identification of a serum lectin from lamprey ( Lampetra japonica ) by chromatographys and
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Target-directed isolation and identification of a serum lectin from lamprey (Lampetra japonica) by chromatographys and MALDI-TOF/TOF HAN Yinglun1, 2†, GOU Meng1, 2†, SONG Xiaoping1, 2, 3†, SONG Tao1, 2†, SHI Biyue1, 2, PANG Yue1, 2, LI Qingwei1, 2* 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
Received 13 March 2016; accepted 29 December 2017 © Chinese Society for Oceanography and Springer-Verlag GmbH Germany, part of Springer Nature 2018
Abstract
A 105-kDa 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 kDa 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 kDa and 35 kDa 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. Key words: Lampetra japonica, lectin, purification, identification, MALDI-TOF/TOF Citation: Han Yinglun, Gou Meng, Song Xiaoping, Song Tao, Shi Biyue, Pang Yue, Li Qingwei. 2018. Target-directed isolation and identification of a serum lectin from lamprey (Lampetra japonica) by chromatographys and MALDI-TOF/TOF. Acta Oceanologica Sinica, 37(5): 113–116, doi: 10.1007/s13131-018-1175-7
1 Introduction Lectins are carbohydrate-binding proteins which have been investigated extensively in recent years for application in medical and immunological research (Ogawa et al., 2011). They have various biological functions from the glycoprotein synthesis to regulation of cell adhesion and the control of protein levels in the animal bloods (Rutishauser and Sachs, 1975). In addition, lectins can serve many important roles in innate immune defense by recognizing carbohydrates (Ourth et al., 2008) and agglutinating red blood cells in vitro (Guo et al., 2013). Agnathans, represented by lamprey and hagfish, are agreed to be the o
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