A unique lectin composing of fibrinogen-like domain from Fenneropenaeus merguiensis contributed in shrimp immune defense and firstly found to mediate encapsulation

In invertebrates, both fibrinogen-related proteins (FREPs) and C-type lectins are acknowledged to act as pattern recognition receptors (PRRs) to participate particularly in an innate immunity. Hereby, a unique C-type lectin designated as FmLFd was isolated from the hemocytes of Fenneropenaeus merguiensis. FmLFd contained one open reading frame which encoding a peptide of 312 amino acid residues and a signal peptide of 18 amino acids. The primary sequence of FmLFd was composed of a fibrinogen-like domain (Fd) with a Ca2+-binding site and possessing specificity to bind N-acetyl glucosamine (G1cNAc). The FtnLFd transcripts were detected mainly in hemocytes of healthy shrimp. The expression of FmLFd was significantly up-regulated upon challenge shrimp with Vibrio parahaemolyticus and Vibrio harveyi which more potent than by white spot syndrome virus (WSSV). The knocking down shrimp with FmLFd double-stranded RNA caused dramatical gene down-regulation. The gene silencing with co-injection of pathogens resulted in reduction of the shrimp survival rate. Recombinant protein of FmLFd (rFmLFd) could agglutinate and bind directly to both Gram-negative and Gram-positive bacteria in a Ca2+-dependent manner and showed the sugar specificity to GlcNAc and bacterial saccharides; peptidoglycan (PGN), lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Recombinant protein of Fd domain (rFd) displayed the lower activity and specificity only to PGN. The binding between recombinant proteins of FmLFd and its domain confirming by ELISA demonstrated that both rFmLFd and rFd could bind to PGN, LPS and LTA with the highest affinity respected to PGN including a less extent of rFd. Besides, rFmLFd but not rFd could bind to WSSV proteins with the highest binding affinity to capsid VP15 and decreasing in order to envelope VP28 and tegument VP39A, respectively. It was presumed that entire molecule of FmLFd exhibited the antimicrobial ability by inhibiting the growth of pathogenic V. parahaemolyticus and this action was not affected by GlcNAc. Otherwise, FmLFd, a lectin containing fibrinogen-like domain, was firstly reported to be capable of promoting encapsulation by hemocytes. Altogether, we concluded that FmLFd belonged to a FREP family indentified by the existence of a conserved fibrinogen-like domain with possessing an ability to bind GlcNAc. It was a new C-type lectin existed in F. merguiensis and might presumably act as a kind of PRRs to participate in the shrimp immune defense towards bacterial and viral pathogens.