Two novel antimicrobial peptides, arasin-likeSp and GRPSp, from the mud crab Scylla paramamosain, exhibit the activity against some crustacean pathogenic bacteria

Antimicrobial peptides (AMPs) are some of the important host molecules required to resist pathogen infection. Two novel AMPs (arasin-likeSp and GRPSp) were identified from the hemocytes of the mud crab, Scylla paramamosain, by analysis of a hemocyte expressed sequence tag library. The deduced open reading frames of the arasin-likeSp and GRPSp cDNAs are 198 and 168 bp, and encode for predicted peptides of 65 and 55 amino acid residues, respectively. The calculated molecular mass of the mature peptides was 4373 and 2995 Da with an estimated isoelectric point (pI) of 11.03 and 9.66, respectively. The mature peptide of arasin-likeSp is predicted to contain an N-terminal region rich in glycine and arginine and a C-terminal region containing four cysteine residues. Its amino acid sequence has an overall sequence identity of 53% to arasin-2 from the spider crab, Hyas araneus. The mature protein of GRPSp contains two cysteine residues at the C-terminus and two glycine-rich repeats (GGYG and GYGG). In healthy crabs, both arasin-likeSp and GRPSp transcript levels were found to be high in the hemocytes and were further increased at 3 h after challenge with the bacterium, Aerococcus viridans. A synthetic arasin-likeSp peptide revealed the antimicrobial activity against both Gram-positive and Gram-negative bacteria including some crustacean pathogens (A. viridans, Vibrio harveyi and V. anguillarum), whilst the synthetic GRPSp peptide exhibited antibacterial activity against some Gram-positive (A. viridans and Micrococcus luteus), but not Gram-negative, bacteria. These results indicate that arasin-likeSp and GRPSp are potentially novel AMPs involved in the immune responses of mud crab, S. paramamosain.