Molecular characterization and phylogenetic analysis of two antimicrobial peptides: Anti-lipopolysaccharide factor and crustin from the brown mud crab, Scylla serrata

AMPs are evolutional weapons, widely used by animals and plants in their innate immune system to fend off invading microbes. The present study reports characterization of a new ALF isoform (Sc-ALF; HQ638024) and the first crustin (Sc-crustin; HQ638025) from the mud crab, Scylla serrata. The full-length cDNA of Sc-ALF consisted of 477 bp with an ORF of 123 amino acids and a putative signal peptide of 26 amino acids. Sc-ALF had a predicted molecular weight (MW) of 11.17 kDa and theoretical isoelectric point (pI) of 9.95. Two highly conserved cysteine residues and putative LPS binding domain were observed in Sc-ALF. Comparison of amino acid sequences with neighbor-joining tree indicated that Sc-ALF shared maximum similarity with ALF of S. paramamosain. Peptide model of Sc-ALF created using SWISS-MODEL server was found to consist of two α-helices crowded against a four-strand β-sheet. The full-length cDNA of Sc-crustin consisted of 433 base pairs with an ORF of 111 amino acids and a putative signal peptide of 21 amino acids. Comparison of amino acid sequences with a neighbor-joining tree revealed that Sc-crustin shared high identity with other known crustins characterized from S. paramamosain, P. trituberculatus, H. araneus, C. maenas and F. chinensis. A whey-acidic-protein domain could be detected at the C-terminus with the characteristic four disulfide core. Sc-crustin had a predicted MW of 10.24 kDa and a pI of 8.76. Peptide model of Sc-crustin created using SWISS-MODEL server indicated a random coiled structure that is with two possible β-sheets but no helices.

► Study reports an ALF isoform and the first crustin sequence from Scylla serrata.
► Sc-ALF has a predicted MW of 11.17 kDa and theoretical pI of 9.95.
► Highly conserved cysteine residues and LPS binding domain were observed in Sc-ALF.
► Sc-crustin shared 81% identity with the crustin from Scylla paramamosain.
► Sc-crustin has a predicted MW of 10.24 kDa and a theoretical pI of 8.