Characterization of a novel short-type peptidoglycan recognition protein from the sea cucumber Apostichopus japonicus.

Abstract

Peptidoglycan recognition proteins (PGRPs) represent a key component of the family of pattern recognition receptors (PRRs). The functional mechanisms of PGRPs in innate immunity are poorly understood. In this study, we identified a novel short-type PGRP, AjPGRP-S2, from the sea cucumber Apostichopus japonicus. Our data showed that AjPGRP-S2 encoded an extracellular protein that possessed a signal peptide, a complete zinc (Zn2+) binding site, and a disulfide bond. A recombinant AjPGRP-S2 (rAjPGRP-S2) lacking the signal peptide was generated and demonstrated to exhibit amidase activity. Tissue expression analysis revealed that AjPGRP-S2 was highly expressed in coelomocytes and tube feet. Immune-responsive analysis indicated that AjPGRP-S2 was able to bind to various pathogen-associated molecular patterns (PAMPs) from bacteria and fungi, as well as to Gram-positive and -negative bacteria, and was majorly induced by DAP-PGN challenge. Basing on RNA-Seq and Pearson's correlation testing, RNA interference, and pull-down analysis, AjPGRP-S2 was found to be involved in transducing immune signals to the complement system and other PRRs, such as fibrinogen, by protein interactions to further recognize and kill pathogens. To respond comprehensively against pathogenic invasion, AjPGRP-S2 may also have the potential in transducing immune signals to key processes, such as cell adhesion, nerve conduction, apoptosis, and transcription by complex pathways that have yet to be elucidated. Our findings not only promote our understanding of the immune-related function and mechanisms of the PGRP family in A. japonicus, but also provide important data that will facilitate the identification of key evolutionary characteristics associated with invertebrate PGRPs.