Characterisation and functional analysis of an L-type lectin from the swimming crab Portunus trituberculatus

Abstract

L-type lectins are involved in glycoprotein secretion and are associated with immune responses. Herein, an L-type lectin was identified in swimming crab (Portunus trituberculatus). The 1347 bp PtLTL cDNA includes a 26 bp 5′-untranslated region (UTR), a 547 bp 3′-UTR with a poly(A) tail, and a 774 bp open reading frame encoding a 257 amino acid protein with a putative 21 residue signalling peptide. The protein includes an L-type lectin carbohydrate recognition domain containing four conserved cysteines. The 714 bp cDNA fragment encoding the mature peptide of PtLTL1 was recombined into pET-21a (+) with a C-terminally hexa-histidine tag fused in-frame and expressed in Escherichia coli Origami (DE3). Recombinant PtLTL1 caused agglutination of all three Gram-positive and Gram-negative bacterial strains tested. In addition, erythrocyte agglutination and LPS-binding activity were observed. PtLTL1 mRNA transcripts were most abundant in P. trituberculatus hepatopancreas and hemocytes, and expression was up-regulated in hemocytes challenged with Vibrio alginolyticus, suggesting PtLTL functions in the immune response against bacterial pathogens.