Antimicrobial peptides (AMPs) are components of innate immunity found in many forms of life. However, there have been no reports of AMPs in sea star (Phylum Echinodermata). Here we report the isolation and characterization of a novel antimicrobial peptide from the coelomic epithelium extract of the sea star Patiria pectinifera. The isolated peptide comprises 38 amino acid residues, is cationic (pI 9.2), has four cysteine residues that form two disulfide bonds (C1-C3 and C2-C4), is amidated at the C-terminus, and is designated P. pectinifera cysteine-rich antimicrobial peptide (PpCrAMP). Synthetic PpCrAMP identical to the native peptide exhibited the most potent antimicrobial activity compared to analogs with different disulfide bond configurations. Expression analysis of PpCrAMP precursor transcripts revealed constitutive expression in the coelomic epithelium and tube feet of P. pectinifera. Analysis of genomic DNA and cDNA encoding the PpCrAMP precursor protein revealed that an intron splits the coding region of the mature peptide into a positively charged N-terminal domain and a C-terminal domain harboring four cysteine residues and a glycine for C-terminal amidation. No significant homology with other known AMPs was observed, while orthologs of PpCrAMP were found in other echinoderm species. These findings indicate that PpCrAMP is the prototype of a family a novel cysteine-rich AMPs that participate in mechanisms of innate immunity in echinoderms. Furthermore, the discovery of PpCrAMP may lead to the identification of related AMPs in vertebrates and protostome invertebrates.