Identification and characterization of a novel ApeC-containing transmembrane protein family in parasitic flatworms

The Apextrin C-terminal (ApeC) domain is widely distributed across various animal clades. Several ApeC domains have been identified as carbohydrate-binding domains, contributing to extracellular adhesion and mucosal immunity. However, the functions and evolutionary history of most ApeC-containing protein (ACP) families remain largely unexplored. In this study, we identified 213 ACPs in flatworms (Platyhelminthes), with each species containing one to two such proteins. All flatworm ACPs belong to a unique transmembrane protein family, characterized by a length of 1700-2200 amino acids and a distinctive domain architecture (SP-[Kringle]_1-2-[ApeC]₇-[EGF]_1-4-TM) unlike any found in other phyla. This ACP is conserved across all major parasitic flatworm lineages, including flukes (Trematoda), tapeworms (Cestoda) and monogeneans (Monogenea), despite their diverse morphologies and habitats. Notably, it is also present in one group of free-living flatworms, the planarians (Tricladida), suggesting that this ACP originated in the free-living ancestor of parasitic flatworms. This ACP contains seven consecutive ApeC domains, an unparalleled number among known animal proteins. Five of these ApeC domains are highly divergent, necessitating the definition of a new domain model (ApeC_Pla; Pfam: PF24148) for accurate classification. Structural predictions indicate that these ApeC domains adopt a conserved three β-sheet structure. Furthermore, transcriptomic analysis revealed that flatworm ACPs, along with several important serpins and proteases, are predominantly expressed in parenchymal cells and feeding organs, suggesting that ACPs serve as novel marker genes for parenchymal tissue and may be involved in cell adhesion, oral immunity, and parasite-host interactions. Taken together, our findings indicate that this flatworm ACP represents a promising target for vaccine development and provides key insights into the physiology of the parenchyma, a unique flatworm tissue that functions as a substitute for a body cavity.