Antimicrobial properties and immune-related gene expression of a C-type lectin isolated from Pinctada fucata martensii

Abstract

C-type lectins are carbohydrate-binding proteins that play important roles in the innate immune response to pathogen infections. Here, multi-step high performance liquid chromatography (HPLC), combined with mass spectrometry (MS), was used to isolate and identify proteins with antibacterial activity from the serum of Pinctada fucata martensii. Using this method, we obtained a novel isoform of C-type lectin (PmCTL-1). PmCTL-1 strongly inhibited gram-positive bacteria. The complete cDNA sequence of PmCTL-1 was 636 bp in length, and encoded a protein 149 amino acids long, containing a typical carbohydrate recognition domain (CRD). A phylogenetic analysis based on a multiple sequence alignment indicated that PmCTL-1 was highly similar to C-type lectins from other mollusks. Fluorescent quantitative real-time PCR analysis showed that PmCTL-1 mRNA was strongly upregulated in the mantle of healthy P.f. martensii, but was expressed only at low levels in the gill, gonad, hepatopancreas, adductor muscle, and hemocytes. PmCTL-1 expression levels in the mantle and hemocytes increased significantly in response to bacterial stimulation. This study provides a valuable framework for further explorations of innate immunity and the immune response in mollusks.