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A novel C-type lectin from crab Eriocheir sinensis functions as pattern recognition receptor enhancing cellular encapsulation

Wang, Leilei, Wang, Lingling, Zhang, Daoxiang, Li, Fengmei, Wang, Mengqiang, Huang, Mengmeng, Zhang, Huan, Song, Linsheng
Fish & shellfish immunology 2013 v.34 no.3 pp. 832-842
crabs, Komagataella pastoris, Gram-positive bacteria, polypeptides, disulfide bonds, Eriocheir sinensis, open reading frames, Vibrio anguillarum, binding proteins, amino acids, hepatopancreas, signal peptide, carbohydrate binding, gene expression regulation, pathogens, Gram-negative bacteria, messenger RNA, complementary DNA, yeasts, encapsulation, immune response, beta-glucans, innate immunity, lectins, hemocytes
C-type lectins are a large family of Ca2+-dependent carbohydrate binding proteins which play crucial roles to recognize and eliminate pathogens in innate immunity. In the present study, a novel C-type lectin was identified from Eriocheir sinensis (designated as EsCTL). The full-length cDNA of EsCTL was of 789 bp with an open reading frame of 468 bp encoding a polypeptide of 156 amino acids with a signal sequence and single carbohydrate-recognition domain (CRD). The potential tertiary structure of the CRD adopted a typical double-loop structure with Ca2+-binding site 2 in the long loop region and two conserved disulfide bridges at the bases of the loops. An EPQ motif to determine carbohydrate binding specificity was identified in the CRD of EsCTL. The mRNA transcripts of EsCTL were mainly detected in hepatopancreas and its relative expression level in hemocytes was significantly up-regulated after the challenges of Vibrio anguillarum (P < 0.05) and Pichia pastoris (P < 0.05). The recombinant EsCTL protein (rEsCTL) could bind different PAMPs, including LPS, PGN, β-glucan, and polyI:C; and also bind various microorganisms including three Gram-positive bacteria, three Gram-negative bacteria and two yeasts. Moreover, rEsCTL could significantly enhance the in vitro encapsulation of crab hemocytes. All these results suggested that EsCTL functioned as an important PRR involved in immune defense against invading pathogen in crab.